Benzimidazole derivatives as bromodomain inhibitors

ABSTRACT

Compounds of formula (I) and salts thereof: 
     
       
         
         
             
             
         
       
     
     wherein R 1 , R 2 , R 3 , R 4  are defined herein. Compounds of formula (I) and salts thereof have been found to inhibit the binding of the BET family of bromodomain proteins to, for example, acetylated lysine residues and thus may have use in therapy, for example in the treatment of autoimmune and inflammatory diseases, such as rheumatoid arthritis; and cancers.

FIELD OF THE INVENTION

The present invention relates to compounds, processes for theirpreparation, compositions containing them, and to their use in thetreatment of various disorders in particular inflammatory and autoimmunediseases, such as rheumatoid arthritis; and cancers.

BACKGROUND TO THE INVENTION

The genomes of eukaryotic organisms are highly organised within thenucleus of the cell. The long strands of duplex DNA are wrapped aroundan octomer of histone proteins (most usually comprising two copies ofhistones H2A, H2B, H3 and H4) to form a nucleosome. This basic unit isthen further compressed by the aggregation and folding of nucleosomes toform a highly condensed chromatin structure. A range of different statesof condensation are possible, and the tightness of this structure variesduring the cell cycle, being most compact during the process of celldivision. Chromatin structure plays a critical role in regulating genetranscription, which cannot occur efficiently from highly condensedchromatin. The chromatin structure is controlled by a series of posttranslational modifications to histone proteins, notably histones H3 andH4, and most commonly within the histone tails which extend beyond thecore nucleosome structure. These modifications include acetylation,methylation, phosphorylation, ubiquitinylation, SUMOylation. Theseepigenetic marks are written and erased by specific enzymes, which placetags on specific residues within the histone tail, thereby forming anepigenetic code, which is then interpreted by the cell to allowregulation of gene expression.

Histone acetylation is most usually associated with the activation ofgene transcription, as the modification relaxes the interaction of theDNA and the histone octomer by changing the electrostatics. In additionto this physical change, specific proteins recognise and bind toacetylated lysine residues within histones to read the epigenetic code.Bromodomains are small (˜110 amino acid) distinct domains withinproteins that bind to acetylated lysine resides commonly but notexclusively in the context of histones. There is a family of around 50proteins known to contain bromodomains, and they have a range offunctions within the cell.

The BET family of bromodomain containing proteins comprises 4 proteins(BRD2, BRD3, BRD4 and BRDT) which contain tandem bromodomains capable ofbinding to two acetylated lysine residues in close proximity, increasingthe specificity of the interaction. Numbering from the N-terminal end ofeach BET protein the tandem bromodomains are typically labelled BindingDomain 1 (BD1) and Binding Domain 2 (BD2) (Chung eta, J Med. Chem.,2011, 54, 3827-3838).

Inhibiting the binding of a BET protein to acetylated lysine residueshas the potential to ameliorate progression of several diseases,including but not limited to, cancer (Dawson M. A. et al, Nature, 2011:478(7370):529-33; Wyce, A. et al, Oncotarget. 2013: 4(12):2419-29),sepsis (Nicodeme E et al, Nature, 2010: 468(7327):1119-23), autoimmuneand inflammatory diseases such as rheumatoid arthritis and multiplesclerosis (Mele D. A. et al, Journal of Experimental Medicine, 2013:210(11):2181-90), heart failure (Anand P. et al, Cell, 2013:154(3):569-82), and lung fibrosis (Tang X et al, Molecular Pharmacology,2013: 83(1):283-293).

There exists a need for chemical compounds which inhibit the activity ofbromodomains, in particular compounds that inhibit the binding of BETproteins to acetylated lysine residues and hence have utility in thetreatment of, for example, autoimmune and inflammatory diseases, andcancers.

SUMMARY OF THE INVENTION

In a first aspect, the present invention is directed to compounds offormula (I) and salts thereof:

whereinR₁ and R₂ are each independently hydrogen or methyl with the provisothat at least one of R₁ and R₂ is methyl;R₃ is C₁₋₆alkyl, C₁₋₆alkoxy, cycloalkyl, heterocycloalkyl, or—CHR₅(CH₂)_(a)R₆;R₄ is attached at the 5 or 6 position of the benzimidazole andrepresents

R₅ is hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, or —(CH₂)_(d)OR₉;R₆ is hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, —(CH₂)_(e)OR₁₀, hydroxyl,—NR₁₁R₁₂, halo, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl,wherein said C₁₋₃alkyl, C₁₋₃alkoxy, —(CH₂)_(e)OR₁₀, cycloalkyl,heterocycloalkyl, aryl or heteroaryl may be optionally substituted withone or two substituents independently selected from the group consistingof C₁₋₃alkyl, C₁₋₃alkoxy, halo, —CH₂OH, —COOH, and —COCH₃;R₇ is hydrogen, C₁₋₆alkyl, —(CH₂)_(g)cycloalkyl,—(CH₂)_(h)heterocycloalkyl, or —CR₁₃R₁₄R₁₅;R₈ is hydrogen, C₁₋₆alkyl, cycloalkyl, heterocycloalkyl, or —CHR₁₆R₁₇wherein said C₁₋₆alkyl is optionally substituted with C₁₋₃alkoxy, andwherein R₁₆ is hydrogen or C₁₋₃alkyl and R₁₇ is cycloalkyl orheterocycloalkyl;R₉, R₁₀, R₁₁, R₁₂, R₁₄, R₁₅, and R₁₈ are each independently hydrogen orC₁₋₃alkyl;R₁₃ is hydrogen, hydroxyl, —CH₂OR₁₈, halo, —COOH, —CONH₂,1H-imidazol-4-yl, —SH, —SeH, C₁₋₃alkyl, C₁₋₃alkoxy, phenyl, or4-hydroxyphenyl, wherein said C₁₋₃alkyl or C₁₋₃alkoxy may be optionallysubstituted with halo or hydroxyl, —NHC(═NH₂)NH₂, —NH₂, —COOH, —CONH₂,or —SCH₃;a is 0, 1, 2 or 3;b is 0 or 1;c is 1, 2 or 3 with the proviso that when b is 1, c is 2 or 3;d and e are each independently 1 or 2; andg and h are each independently 0, 1 or 2.

Compounds of formula (I) and salts thereof have been found to inhibitthe binding of bromodomain proteins; in particular, the binding of theBET family of bromodomain proteins to, for example, acetylated lysineresidues. Compounds of formula (I), or pharmaceutically acceptable saltsthereof, may thus have use in therapy, for example in the treatment ofautoimmune and inflammatory diseases, such as rheumatoid arthritis; andcancers.

The present invention is further directed to methods of treatment ofautoimmune and inflammatory diseases and cancers through inhibition ofthe function of bromodomain proteins, for example members of the BETfamily of bromodomain proteins, which comprises administering to asubject in need thereof, a therapeutically effective amount of acompound of formula (I), or a pharmaceutically acceptable salt thereof.

In a further aspect, the present invention is directed to pharmaceuticalcompositions comprising a compound of formula (I), or a pharmaceuticallyacceptable salt thereof, and one or more pharmaceutically acceptableexcipients.

DETAILED DESCRIPTION OF THE INVENTION Definitions

As used herein, the term “bromodomain” refers to evolutionary andstructurally conserved modules (approximately 110 amino acids in length)that bind acetylatedlysine residues, such as those on the N-terminaltails of histones. They are protein domains that are found as part ofmuch larger bromodomain containing proteins (BCPs), many of which haveroles in regulating gene transcription and/or chromatin remodelling. Thehuman genome encodes for at least 57 bromodomains.

As used herein, the term “BET” refers to the bromodomain andextraterminal domain family of bromodomain containing proteins whichinclude BRD2, BRD3, BRD4 and BRDT.

As used herein, the term “BET inhibitor” refers to a compound that iscapable of inhibiting the binding of one or more BET family bromodomaincontaining proteins (e.g. BRD2, BRD3, BRD4 or BRDT) to, for example,acetylated lysine residues.

As used herein, the term “alkyl” refers to a saturated hydrocarbonchain, straight or branched, having the specified number of carbonatoms. For example, C₁₋₆ alkyl refers to an alkyl group having from 1 to6 carbon atoms. Unless otherwise stated, alkyl groups are unsubstituted.The term “alkyl” includes, but is not limited to, methyl, ethyl, propyl(n-propyl and isopropyl), butyl (n-butyl, sec-butyl, isobutyl andtert-butyl), pentyl, and hexyl.

As used herein, the term “alkoxy” refers to an —O-alkyl group wherein“alkyl” is defined above.

As used herein, the term “cycloalkyl” refers to a saturated, monocyclic,hydrocarbon ring having 3 (cyclopropyl), 4 (cyclobutyl), 5(cyclopentyl), 6 (cyclohexyl) or 7 (cycloheptyl) carbon atoms.

As used herein, the term “heterocycloalkyl” refers to a saturated orunsaturated 3 to 7 membered monocyclic or bicyclic ring, which mustcontain 1 or 2 non-carbon atoms, which are selected from nitrogen,oxygen, and sulfur. Heterocycloalkyl groups may contain one or moreC(O), S(O) or SO₂ groups. However, heterocycloalkyl groups are notaromatic. Heterocycloalkyl groups containing more than one heteroatommay contain different heteroatoms. “5 or 6 membered heterocycloalkyl”refers to a saturated or unsaturated 5 or 6 membered monocyclic ring,which must contain 1 or 2 non-carbon atoms, which are selected fromnitrogen, oxygen, and sulfur. Heterocycloalkyl includes, but is notlimited to, pyrrolidine, piperidine, piperazine, oxetane,tetrahydrofuran, tetrahydro-2H-pyran, morpholine, morpholine-3-one,piperidin-2-one, pyrimidine-2,4(1H,3H)-dione, thiomorpholine, andthiomorpholine 1,1-dioxide.

As used herein, the term “aryl” refers to a monocyclic or bicyclic,hydrocarbon, aromatic radical. Aryl includes, for example, phenyl andnaphthyl.

As used herein, the term “heteroaryl” refers to a monocyclic orbicyclic, aromatic radical containing one or more heteroatoms selectedfrom S, N and O. Illustrative examples of heteroaryl useful in thepresent invention include, but are not limited to, furanyl, thienyl,pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl,oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl,pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, benzofuranyl,isobenzofuryl, 2,3-dihydrobenzofuryl, 1,3-benzodioxolyl,dihydrobenzodioxinyl, benzothienyl, indolizinyl, indolyl, isoindolyl,dihydroindolyl, benzimidazolyl, dihydrobenzimidazolyl, benzoxazolyl,dihydrobenzoxazolyl, benzthiazolyl, benzoisothiazolyl,dihydrobenzoisothiazolyl, indazolyl, imidazopyridinyl,pyrazolopyridinyl, benzotriazolyl, triazolopyridinyl, purinyl,quinolinyl, tetrahydroquinolinyl, isoquinolinyl,tetrahydroisoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl,quinazolinyl, 1,5-naphthyridinyl, 1,6-naphthyridinyl,1,7-naphthyridinyl, 1,8-naphthyridinyl, and pteridinyl.

As used herein, the phrase “optionally substituted” indicates that agroup may be unsubstituted or substituted with one or more substituentsas defined herein. “Substituted” in reference to a group indicates thata hydrogen atom attached to a member atom within a group is replaced byone of the defined substituents.

As used herein, the term “pharmaceutically acceptable salts” refers tosalts that retain the desired biological activity of the subjectcompound and exhibit minimal undesired toxicological effects. Thesepharmaceutically-acceptable salts may be prepared in situ during thefinal isolation and purification of the compound, or by separatelyreacting the purified compound in its free acid or free base form with asuitable base or acid, respectively. Furthermore,pharmaceutically-acceptable salts of the compound of formula (I) may beprepared during further processing of the free acid or base form, forexample in situ during manufacture into a pharmaceutical formulation.

As used herein, the phrase “attached at the 5 or 6 position of thebenzimidazole” refers to attachment of the specified substituent at the5 or 6 position as denoted in the structure below:

As used herein, the term “treatment” refers to prophylaxis of thecondition, ameliorating or stabilising the specified condition, reducingor eliminating the symptoms of the condition, slowing or eliminating theprogression of the condition, and preventing or delaying reoccurrence ofthe condition in a previously afflicted patient or subject. In oneembodiment, treatment refers to ameliorating or stabilising a specifiedcondition, reducing or eliminating the symptoms of the condition, orslowing or eliminating the progression of the condition.

As used herein, the term “therapeutically effective amount” refers tothe quantity of a compound of formula (I), or a pharmaceuticallyacceptable salt thereof, which will elicit the desired biologicalresponse in an animal or human body.

As used herein, the term “subject” refers to an animal or human body.

It is to be understood that references herein to “compound(s) of theinvention” mean a compound of formula (I) as the free base, or as asalt, for example a pharmaceutically acceptable salt.

Statement of the Invention

In a first aspect, the present invention provides compounds of formula(I), or salts thereof:

whereinR₁ and R₂ are each independently hydrogen or methyl with the provisothat at least one of R₁ and R₂ is methyl;R₃ is C₁₋₆alkyl, C₁₋₆alkoxy, cycloalkyl, heterocycloalkyl, or—CHR₅(CH₂)_(a)R₆;R₄ is attached at the 5 or 6 position of the benzimidazole andrepresents

R₅ is hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, or —(CH₂)_(d)OR₉;R₆ is hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, —(CH₂)_(e)OR₁₀, hydroxyl,—NR₁₁R₁₂, halo, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl,wherein said C₁₋₃alkyl, C₁₋₃alkoxy, —(CH₂)_(e)OR₁₀, cycloalkyl,heterocycloalkyl, aryl or heteroaryl may be optionally substituted withone or two substituents independently selected from the group consistingof C₁₋₃alkyl, C₁₋₃alkoxy, halo, —CH₂OH, —COOH, and —COCH₃;R₇ is hydrogen, C₁₋₆alkyl, —(CH₂)_(g)cycloalkyl,—(CH₂)_(h)heterocycloalkyl, or —CR₁₃R₁₄R₁₅;R₈ is hydrogen, C₁₋₆alkyl, cycloalkyl, heterocycloalkyl, or —CHR₁₆R₁₇wherein said C₁₋₆alkyl is optionally substituted with C₁₋₃alkoxy, andwherein R₁₆ is hydrogen or C₁₋₃alkyl and R₁₇ is cycloalkyl orheterocycloalkyl;R₉, R₁₀, R₁₁, R₁₂, R₁₄, R₁₅, and R₁₈ are each independently hydrogen orC₁₋₃alkyl;R₁₃ is hydrogen, hydroxyl, CH₂OR₁₈, halo, —COOH, —CONH₂,1H-imidazol-4-yl, —SH, —SeH, C₁₋₃alkyl, C₁₋₃alkoxy, phenyl, or4-hydroxyphenyl, wherein said C₁₋₃alkyl or C₁₋₃alkoxy may be optionallysubstituted with halo or hydroxyl, —NHC(═NH₂)NH₂, —NH₂, —COOH, —CONH₂,or —SCH₃;a is 0, 1, 2 or 3;b is 0 or 1;c is 1, 2 or 3 with the proviso that when b is 1, c is 2 or 3;d and e are each independently 1 or 2; andg and h are each independently 0, 1 or 2.

In a further aspect of the present invention, R₁ and R₂ are each methyl.

In a further embodiment, R₃ represents 5 or 6 membered heterocycloalkylor the group —CHR₅R₆ wherein R₅ represents hydrogen or C₁₋₃alkyl and R₆represents 5 or 6 membered heterocycloalkyl, further wherein saidheterocycloalkyl may be optionally substituted with C₁₋₃alkyl, orC(O)CH₃.

In a further embodiment, R₆ is selected from the group consisting of:

In a further embodiment of the present invention, R₆ is:

In a further embodiment of the present invention, R₃ is the group—CHR₅R₆ wherein R₅ is hydrogen and R₆ is

In a further embodiment of the present invention, R₃ is selected fromthe group consisting of:

wherein Ra represent hydrogen or C₁₋₃alkyl.

In a further aspect of the present invention, R₅ is hydrogen.

In a further aspect of the present invention, R₃ is the group—CHR₅(CH₂)_(a)R₆, and further wherein R₅ is —(CH₂)_(d)OR₉, a is 0 and R₆is —(CH₂)_(e)OR₁₀.

In a further aspect of the present invention, R₅ and R₆ each represent—CH₂OCH₃.

In a further aspect of the present invention, R₄ is attached at the 5position of the benzimidazole.

In a further aspect of the present invention, b is 0.

In a further aspect of the present invention, c is 1.

In a further aspect of the present invention, R₄ represents:

In a further aspect of the present invention, R₇ is hydrogen, methyl,isopropyl, sec-butyl, isobutyl, —CH₂-phenyl, —CH₂-4-hydroxyphenyl,—CH₂OH, —CH(CH₃)OH, —CH₂SH, —CH₂SeH, —(CH₂)₂SCH₃, —CH₂COOH, —(CH₂)₂COOH,—CH₂CONH₂, —(CH₂)₂CONH₂, —(CH₂)₄NH₂, —(CH₂)₃NHC(═NH₂)NH₂, or—CH₂-1H-imidazol-4-yl.

In a further aspect of the present invention, R₇ is isopropyl,sec-butyl, or —CH(CH₃)OH.

In a further aspect of the present invention, R₇ is —CH(CH₃)OH.

In a further aspect of the present invention, R₇ is (R)-1-hydroxyethyl.

In a further aspect of the present invention, R is hydrogen.

In a further aspect of the present invention, R is C₁₋₆alkyl orcycloalkyl.

In a further aspect of the present invention, R is isopropyl, isobutylor cyclopentyl.

In a further aspect of the present invention, R is isopropyl.

The present invention covers all combinations of substituent groupsreferred to herein above.

In a further aspect, the present invention provides a compound offormula (Ia):

whereinR₃ is cycloalkyl, hetercycloalkyl, or the group —CHR₅(CH₂)_(a)R₆;R₅ is hydrogen, C₁₋₃alkyl, or C₁₋₃alkoxy;R₆ is cycloalkyl or heterocycloalkyl, wherein said cycloalkyl orheterocycloalkyl may be optionally substituted with one or twosubstituents selected from the group consisting of C₁₋₃alkyl,C₁₋₃alkoxy, halo, —CH₂OH, —COOH, and —COCH₃;R₇ is hydrogen, C₁₋₆alkyl, —(CH₂)_(g)cycloalkyl,—(CH₂)_(h)heterocycloalkyl, or —CR₁₃R₁₄R₁₅;R₈ is hydrogen, C₁₋₆alkyl, cycloalkyl or heterocycloalkyl, wherein saidC₁₋₆alkyl is optionally substituted with C₁₋₃alkoxy;R₁₃ is hydrogen, hydroxyl, —CH₂OR₁₈, halo, —COOH, —CONH₂,1H-imidazol-4-yl, —SH, —SeH, C₁₋₃alkyl, C₁₋₃alkoxy, phenyl, or4-hydroxyphenyl wherein said C₁₋₃alkyl or C₁₋₃alkoxy may be optionallysubstituted with halo, hydroxyl, —NHC(═NH₂)NH₂, —NH₂, —COOH, —CONH₂, or—SCH₃;R₁₄, R₁₅ and R₁₈ are each independently hydrogen or C₁₋₃alkyl; anda, g and h are each independently 0, 1 or 2.

In a further aspect, the present invention provides a compound offormula (Ib):

whereinR₃ is cycloalkyl, hetercycloalkyl, or the group —CHR₅(CH₂)_(a)R₆;R₅ is hydrogen, C₁₋₃alkyl, or C₁₋₃alkoxy;R₆ is cycloalkyl or heterocycloalkyl, wherein said cycloalkyl orheterocycloalkyl may be optionally substituted with one or twosubstituents selected from the group consisting of C₁₋₃alkyl,C₁₋₃alkoxy, halo, —CH₂OH, —COOH, and —COCH₃;R₇ is hydrogen, C₁₋₆alkyl, —(CH₂)_(g)cycloalkyl,—(CH₂)_(h)heterocycloalkyl, or —CR₁₃R₁₄R₁₅;R₈ is hydrogen, C₁₋₆alkyl, cycloalkyl, or heterocycloalkyl, wherein saidC₁₋₆alkyl is optionally substituted with C₁₋₃alkoxy;R₁₃ is hydrogen, hydroxyl, —CH₂OR₁₈, halo, —COOH, —CONH₂,1H-imidazol-4-yl, —SH, —SeH, C₁₋₃alkyl, C₁₋₃alkoxy, phenyl, or4-hydroxyphenyl wherein said C₁₋₃alkyl or C₁₋₃alkoxy may be optionallysubstituted with halo, hydroxyl, —NHC(═NH₂)NH₂, —NH₂, —COOH, —CONH₂, or—SCH₃;R₁₄, R₁₅ and R₁₈ are each independently hydrogen or C₁₋₃alkyl; anda, g and h are each independently 0, 1 or 2.

In a further aspect, the present invention provides a compound offormula (Ia):

whereinR₃ is C₁₋₆alkyl, C₁₋₆alkoxy, cycloalkyl, heterocycloalkyl, or—CHR₅(CH₂)_(a)R₆;R₅ is hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, or —(CH₂)_(d)OR₉;R₆ is hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, —(CH₂)_(e)OR₁₀, hydroxyl,—NR₁₁R₁₂, halo, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl,wherein said C₁₋₃alkyl, C₁₋₃alkoxy, —(CH₂)_(e)OR₁₀, cycloalkyl,heterocycloalkyl, aryl or heteroaryl may be optionally substituted withone or two substituents independently selected from the group consistingof C₁₋₃alkyl, C₁₋₃alkoxy, halo, —CH₂OH, —COOH, and —COCH₃;R₇ is C₁₋₆alkyl, —CH₂OH, —CH₂OCH₃, —(CH₂)₂OCH₃, —CH(CH₃)OH, or—C(CH₃)₂OH;R₈ is hydrogen, C₁₋₆alkyl, cycloalkyl, or heterocycloalkyl, wherein saidC₁₋₆alkyl is optionally substituted with C₁₋₃alkoxy;R₉, R₁₀, R₁₁, and R₁₂ are each independently hydrogen or C₁₋₃alkyl;a is 0, 1 or 2; andd and e are each independently 1 or 2.

In a further aspect, the present invention provides a compound offormula (Ib):

whereinR₃ is C₁₋₆alkyl, C₁₋₆alkoxy, cycloalkyl, heterocycloalkyl, or—CHR₅(CH₂)_(a)R₆;R₅ is hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, or —(CH₂)_(d)OR₉;R₆ is hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, —(CH₂)_(e)OR₁₀, hydroxyl,—NR₁₁R₁₂, halo, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl,wherein said C₁₋₃alkyl, C₁₋₃alkoxy, —(CH₂)_(e)OR₁₀, cycloalkyl,heterocycloalkyl, aryl or heteroaryl may be optionally substituted withone or two substituents independently selected from the group consistingof C₁₋₃alkyl, C₁₋₃alkoxy, halo, —CH₂OH, —COOH, and —COCH₃;R₇ is C₁₋₆alkyl, —CH₂OH, —CH₂OCH₃, —(CH₂)₂OCH₃, —CH(CH₃)OH, or—C(CH₃)₂OH;R₈ is hydrogen, C₁₋₆alkyl, cycloalkyl, or heterocycloalkyl, wherein saidC₁₋₆alkyl is optionally substituted with C₁₋₃alkoxy;R₉, R₁₀, R₁₁, and R₁₂ are each independently hydrogen or C₁₋₃alkyl;a is 0, 1 or 2; andd and e are each independently 1 or 2.

In a further aspect, the present invention provides a compound offormula (Ia):

whereinR₃ is cycloalkyl, hetercycloalkyl, or —CHR₅(CH₂)_(a)R₆;R₅ is hydrogen or C₁₋₃alkyl;R₆ is cycloalkyl or heterocycloalkyl, wherein said cycloalkyl orheterocycloalkyl may be optionally substituted with one or twosubstituents independently selected from the group consisting ofC₁₋₃alkyl, C₁₋₃alkoxy, halo, —CH₂OH, —COOH, and —COCH₃;R₇ is C₁₋₆alkyl, —CH₂OH, —CH₂OCH₃, —(CH₂)₂OCH₃, —CH(CH₃)OH, or—C(CH₃)₂OH;R₈ is hydrogen, C₁₋₆alkyl, cycloalkyl, or heterocycloalkyl, wherein saidC₁₋₆alkyl is optionally substituted with C₁₋₃alkoxy; anda is 0, 1 or 2.

In a further aspect, the present invention provides a compound offormula (Ib):

whereinR₃ is cycloalkyl, hetercycloalkyl, or —CHR₅(CH₂)_(a)R₆;R₅ is hydrogen or C₁₋₃alkyl;R₆ is cycloalkyl or heterocycloalkyl, wherein said cycloalkyl orheterocycloalkyl may be optionally substituted with one or twosubstituents independently selected from the group consisting ofC₁₋₃alkyl, C₁₋₃alkoxy, halo, —CH₂OH, —COOH, and —COCH₃;R₇ is C₁₋₆alkyl, —CH₂OH, —CH₂OCH₃, —(CH₂)₂OCH₃, —CH(CH₃)OH, or—C(CH₃)₂OH;R₈ is hydrogen, C₁₋₆alkyl, cycloalkyl, or heterocycloalkyl, wherein saidC₁₋₆alkyl is optionally substituted with C₁₋₃alkoxy; anda is 0, 1 or 2.

In a further aspect, the present invention provides a compound offormula (Ia):

wherein R₃ is selected from the group consisting of:

wherein Ra is hydrogen or C₁₋₃alkyl;R₇ is isopropyl, sec-butyl, or —CH(CH₃)OH; andR₈ is isopropyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl,cyclopentyl, or —CH₂OCH₃.

In a further aspect, the present invention provides a compound offormula (Ib):

wherein R₃ is selected from the group consisting of:

wherein Ra is hydrogen or C₁₋₃alkyl;R₇ is isopropyl, sec-butyl, or —CH(CH₃)OH; andR₈ is isopropyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl,cyclopentyl, or —CH₂OCH₃.

In a further aspect, the present invention provides a compound offormula (Ic):

wherein R₆ is selected from the group consisting of:

R₇ is isopropyl, sec-butyl, or —CH(CH₃)OH; andR₈ is hydrogen, C₃₋₆alkyl or cycloalkyl.

In a further aspect, the present invention provides a compound offormula (Id):

wherein R₆ is selected from the group consisting of:

R₇ is isopropyl, sec-butyl, or —CH(CH₃)OH; andR₈ is hydrogen, C₃₋₆alkyl or cycloalkyl.

In a further aspect, the present invention provides a compound offormula (Ic):

wherein R₆ is selected from the group consisting of:

R₇ is isopropyl, sec-butyl, or —CH(CH₃)OH; andR₈ is isopropyl, isobutyl or cyclopentyl.

In a further aspect, the present invention provides a compound offormula (Id):

wherein R₆ is selected from the group consisting of:

R₇ is isopropyl, sec-butyl, or —CH(CH₃)OH; andR₈ is isopropyl, isobutyl or cyclopentyl.

In a further embodiment, the present invention is directed to compoundsof formula (I) and salts thereof:

whereinR₁ and R₂ are each independently hydrogen or methyl with the provisothat at least one of R₁ and R₂ is methyl;R₃ is C₁₋₆alkyl, C₁₋₆alkoxy, cycloalkyl, heterocycloalkyl, or—CHR₅(CH₂)_(a)R₆;R₄ is attached at the 5 or 6 position of the benzimidazole andrepresents

R₅ is hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, or —(CH₂)_(d)OR₉;R₆ is hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, —(CH₂)_(e)OR₁₀, hydroxyl,—NR₁₁R₁₂, halo, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl,wherein said C₁₋₃alkyl, C₁₋₃alkoxy, —(CH₂)_(e)OR₁₀, cycloalkyl,heterocycloalkyl, aryl or heteroaryl may be optionally substituted withone or two substituents independently selected from the group consistingof C₁₋₃alkyl, C₁₋₃alkoxy, halo, —CH₂OH, —COOH, and —COCH₃;R₇ is hydrogen, C₁₋₆alkyl, —(CH₂)_(g)cycloalkyl,—(CH₂)_(h)heterocycloalkyl, or —CR₁₃R₁₄R₁₅;R₈ is C₁₋₆alkyl, cycloalkyl, heterocycloalkyl, or —CHR₁₆R₁₇ wherein saidC₁₋₆alkyl is optionally substituted with C₁₋₃alkoxy, and wherein R₁₆ ishydrogen or C₁₋₃alkyl and R₁₇ is cycloalkyl or heterocycloalkyl;R₉, R₁₀, R₁₁, R₁₂, R₁₄, R₁₅, and R₁₈ are each independently hydrogen orC₁₋₃alkyl;R₁₃ is hydrogen, hydroxyl, —CH₂OR₁₈, halo, —COOH, —CONH₂,1H-imidazol-4-yl, —SH, —SeH, C₁₋₃alkyl, C₁₋₃alkoxy, phenyl, or4-hydroxyphenyl, wherein said C₁₋₃alkyl or C₁₋₃alkoxy may be optionallysubstituted with halo or hydroxyl, —NHC(═NH₂)NH₂, —NH₂, —COOH, —CONH₂,or —SCH₃;a is 0, 1, 2 or 3;b is 0 or 1;c is 1, 2 or 3 with the proviso that when b is 1, c is 2 or 3;d and e are each independently 1 or 2; andg and h are each independently 0, 1 or 2.

Specific examples of compounds of formula (I) are:

-   tert-butyl    (2S,3R)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-6-yl)methyl]amino}-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3S)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl)methyl]amino}-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl)methyl]amino}-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-3-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   cyclopentyl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]propanoate;-   propan-2-yl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-6-yl)methyl]amino}-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   cyclobutyl    (2S,3R)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl)methyl]amino}-3-hydroxybutanoate;-   propan-2-yl    (2S)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl)methyl]amino}-3-methylbutanoate;-   cyclopentyl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-methoxypropanoate;-   2,2-dimethylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   cyclobutyl    (2S,3R)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-hydroxybutanoate;-   cyclopentyl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]oxy}ethyl)amino]propanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-3-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-{[(1-{[(3S)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-6-yl)methyl]amino}-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-yl    methyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-yl    methyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   cyclobutyl    (2R,3S)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   (3S)-oxolan-3-yl    (2S,3R)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-hydroxybutanoate;-   cyclopentyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   (2S)-butan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   cyclopentyl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]oxy}ethyl)amino]-3-methoxypropanoate;-   (3S)-oxolan-3-yl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]oxy}ethyl)amino]-3-methyl    butanoate;-   (2S)-1-methoxypropan-2-yl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]oxy}ethyl)amino]-3-methylbutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2S)-oxan-2-ylmethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-2-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-{[(1-{[(3S)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-6-yl)methyl]amino}-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-2-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2R)-oxan-2-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2R)-oxan-2-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2R)-oxan-2-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2R)-oxan-2-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2R)-oxan-2-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   cyclobutyl    (2R,3S)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(1R)-1-(oxan-4-yl)ethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(1R)-1-(oxan-4-yl)ethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({2-[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]ethyl}amino)-3-hydroxybutanoate;-   2,2-dimethylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   cyclopentyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-2-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]oxy}ethyl)amino]-3-hydroxybutanoate;-   2-hydroxy-2-methyl propyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-2-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-2-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-{[(1-{[(3S)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-6-yl)methyl]amino}-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2R)-oxan-2-yl    methyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2S)-oxan-2-yl    methyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2S)-oxan-2-yl    methyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   cyclobutyl    (2S,3R)-2-({[1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   cyclobutyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-{[(1-{[(3S)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl)methyl]amino}-3-hydroxybutanoate;-   cyclobutyl    (2S,3R)-2-{[(1-{[(3S)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl)methyl]amino}-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-{[(1-{[(3S)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl)methyl]amino}-3-hydroxybutanoate;-   propan-2-yl    (2S)-4-chloro-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)butanoate;-   cyclopentyl    (2S)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-4-methylpentanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   (3S)-oxolan-3-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-{[(2R)-4-methylmorpholin-2-yl]methyl}-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   (3S)-oxolan-3-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   cyclobutyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2S)-oxan-2-ylmethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3S)-piperidin-3-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   (3S)-oxolan-3-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-2-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   (3S)-oxolan-3-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2R)-oxan-2-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   (3S)-oxolan-3-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2R)-oxan-2-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[1-(oxan-4-yl)ethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({2-[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]ethyl}amino)-3-hydroxybutanoate;-   cyclopentyl    (2S)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2S)-1-methoxypropan-2-yl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-methoxypropanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3S)-piperidin-3-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2S)-1-methoxypropan-2-yl]-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-methoxypropanoate;-   cyclopentyl    (2S)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxypropanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[1-(oxan-4-yl)ethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3S)-oxolan-3-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3S)-oxolan-3-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(4-methylmorpholin-2-yl)methyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]oxy}ethyl)amino]-3-hydroxybutanoate;-   (3S)-oxolan-3-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-methoxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2S)-1-methoxypropan-2-yl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-3-hydroxy-2-({[2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)butanoate;-   (3S)-oxolan-3-yl    (2S,3S)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2S)-1-methoxypropan-2-yl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2S)-1-methoxypropan-2-yl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(2S)-1-methoxypropan-2-yl]-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxolan-3-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   cyclobutyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxolan-3-ylmethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxolan-3-ylmethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxolan-3-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;    and-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate,    or salts thereof.

In a further embodiment, the present invention provides a compound, or asalt thereof, which is selected from the group consisting of:

-   tert-butyl    (2S,3R)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-6-yl)methyl]amino}-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   2-methylpropyl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3S)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   tert-butyl    (2S,3R)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl)methyl]amino}-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl)methyl]amino}-3-hydroxybutanoate;-   2-methylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-3-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   cyclopentyl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]propanoate;-   propan-2-yl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-6-yl)methyl]amino}-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   cyclobutyl    (2S,3R)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl)methyl]amino}-3-hydroxybutanoate;-   propan-2-yl    (2S)-2-{[(1-{[(3R)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-5-yl)methyl]amino}-3-methylbutanoate;-   cyclopentyl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-methoxypropanoate;-   2,2-dimethylpropyl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   cyclobutyl    (2S,3R)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-hydroxybutanoate;-   cyclopentyl    (2S)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-6-yl]oxy}ethyl)amino]propanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-3-ylmethyl)-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-{[(1-{[(3S)-1-acetylpiperidin-3-yl]methyl}-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-1,3-benzodiazol-6-yl)methyl]amino}-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;-   propan-2-yl    (2S,3R)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-5-yl]methyl}amino)-3-hydroxybutanoate;-   cyclobutyl    (2R,3S)-2-({[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-[(3R)-oxan-3-ylmethyl]-1H-1,3-benzodiazol-6-yl]methyl}amino)-3-hydroxybutanoate;    and-   (3S)-oxolan-3-yl    (2S,3R)-2-[(2-{[2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxan-4-ylmethyl)-1H-1,3-benzodiazol-5-yl]oxy}ethyl)amino]-3-hydroxybutanoate.

In a further embodiment, the present invention provides a compound whichis (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,of formula:

or a salt thereof.

In a further embodiment, the present invention provides a compound whichis(2S,3R)-2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoicacid, of formula:

or a salt thereof.

In a further embodiment, the present invention provides a compound whichis (2S,3R)-Isopropyl2-(((1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,of formula:

or a salt thereof.

In a further embodiment, the present invention provides a compound whichis(2S,3R)-2-(((1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoicacid, of formula:

or a salt thereof.

In a further embodiment of the present invention, a compound of formula(I) is in the form of a free base. In one embodiment, the compound offormula (I) in the form of a free base is any one of the compounds ofExamples 1 to 324.

Salts of the compounds of formula (I) include pharmaceuticallyacceptable salts and salts which may not be pharmaceutically acceptablebut may be useful in the preparation of compounds of formula (I) andpharmaceutically acceptable salts thereof.

In one embodiment of the present invention, a compound of formula (I) isin the form of a pharmaceutically acceptable salt. In one embodiment,the compound of any of Example 1 to 324 is in the form of apharmaceutically acceptable salt.

Compounds of formula (I) may contain an acidic or basic functional groupand, thus, the skilled artisan will appreciate that pharmaceuticallyacceptable salts of the compounds of formula (I) may be prepared.Pharmaceutically acceptable salts of compounds of the invention maypossess, for example, improved stability, solubility, and/orcrystallinity, facilitating development as a medicine.

Compounds of formula (I) may contain a basic functional group and may becapable of forming pharmaceutically acceptable acid addition salts bytreatment with an suitable acid (inorganic or organic acid).Representative pharmaceutically acceptable acid addition salts includehydrochloride, hydrobromide, nitrate, sulfate, bisulfate, sulfamate,phosphate, acetate, hydroxyacetate, phenylacetate, propionate, butyrate,isobutyrate, valerate, maleate, hydroxymaleate, acrylate, fumarate,maleate, tartrate, citrate, salicylate, p-aminosalicyclate, glycollate,lactate, heptanoate, phthalate, oxalate, succinate, benzoate,o-acetoxybenzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate,hydroxybenzoate, methoxybenzoate, naphthoate, hydroxynaphthoate,mandelate, tannate, formate, stearate, ascorbate, palmitate, oleate,pyruvate, pamoate, malonate, laurate, glutarate, glutamate, estolate,methanesulfonate (mesylate), ethanesulfonate (esylate),2-hydroxyethanesulfonate, benzenesulfonate (besylate),p-aminobenzenesulfonate, p-toluenesulfonate (tosylate), andnapthalene-2-sulfonate. In another embodiment, the pharmaceuticallyacceptable salt is the 1,2-ethanedisulphonic acid (edisylate) salt.

Compounds of formula (I) may contain an acidic functional group andsuitable pharmaceutically-acceptable salts include salts of such acidicfunctional groups. Representative salts include pharmaceuticallyacceptable metal salts such as sodium, potassium, lithium, calcium,magnesium, aluminum, and zinc salts; pharmaceutically acceptable organicprimary, secondary, and tertiary amines including aliphatic amines,aromatic amines, aliphatic diamines, and hydroxy alkylamines such asmethylamine, ethylamine, 2-hydroxyethylamine, diethylamine, TEA,ethylenediamine, ethanolamine, diethanolamine, and cyclohexylamine.

In one embodiment, there is provided a compound which is the1,2-ethanedisulphonic acid salt of (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,of formula:

In one embodiment, there is provided a crystalline solid state form of(2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,1,2-ethanedisulphonic acid salt.

In a further embodiment, there is provided a crystalline solid stateform of (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,1,2-ethanedisulphonic acid salt characterised by an X-ray powderdiffraction (XRPD) pattern having significant diffraction peaks at 26values shown in Table 1.

TABLE 1 XRPD peak table for the 1,2-ethanedisulphonic acid salt of(2S,3R)-isopropyl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate Position/°2θ d-spacing [Å] 5.4 16.5 8.810.1 9.9 8.9 11.0 8.1 11.6 7.7 13.5 6.6 13.8 6.4 15.0 5.9 15.7 5.6 16.05.6 16.9 5.2 18.0 4.9 18.4 4.8 18.6 4.8 19.2 4.6 19.4 4.6 19.8 4.5 20.44.3 20.9 4.3 21.1 4.2 21.3 4.2 22.0 4.0 22.4 4.0 22.9 3.9 23.4 3.8 24.93.6 25.1 3.5 25.7 3.5 25.9 3.4 26.4 3.4 26.7 3.3 26.9 3.3 27.5 3.2 28.13.2 28.5 3.1 30.2 3.0 32.2 2.8 32.9 2.7 33.2 2.7 33.6 2.7 34.3 2.6 34.82.6 35.4 2.5 35.7 2.5 36.4 2.5 37.8 2.4 38.3 2.4 38.7 2.3 39.1 2.3

In a further embodiment, there is provided a crystalline solid stateform of (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,1,2-ethanedisulphonic acid salt characterised by an X-ray powderdiffraction (XRPD) pattern having significant diffraction peaks at 28values, ±0.1° 2θ experimental error, of 5.4, 8.8, 9.9, 11.6, 13.8, 16.9,18.0, 16.6, 19.1, 19.4, 19.8, 20.4, 20.9, 21.3, 22.0, 22.4, 22.9, 23.4,24.9, and 25.1 degrees.

For a review on suitable salts see Berge et al., J. Pharm. Sci., 66:1-19(1977). The invention includes within its scope all possiblestoichiometric and non-stoichiometric forms of the salts of thecompounds of formula (I).

Salts may be formed using techniques well-known in the art, for exampleby precipitation from solution followed by filtration, or by evaporationof the solvent.

It will be appreciated that many organic compounds can form complexeswith solvents in which they are reacted or from which they areprecipitated or crystallised. These complexes are known as “solvates”.For example, a complex with water is known as a “hydrate”. Solvents withhigh boiling points and/or solvents with a high propensity to formhydrogen bonds such as water, ethanol, iso-propyl alcohol, and N-methylpyrrolidinone may be used to form solvates. Methods for theidentification of solvated include, but are not limited to, NMR andmicroanalysis. Compounds of formula (I), or salts thereof, may exist issolvated and unsolvated form.

Certain of the compounds of the invention may exist in tautomeric forms.It will be understood that the present invention encompasses all of thetautomers of the compounds of the invention whether as individualtautomers or as mixtures thereof.

The compounds of the invention may be in crystalline or amorphous form.The most thermodynamically stable crystalline form of a compound of theinvention is of particular interest.

Crystalline forms of compounds of the invention may be characterised anddifferentiated using a number of conventional analytical techniques,including, but not limited to, X-ray powder diffraction (XRPD), infraredspectroscopy (IR), Raman spectroscopy, differential scanning calorimetry(DSC), thermogravimetric analysis (TGA) and solid-state nuclear magneticresonance (ssNMR).

The present invention also includes all suitable isotopic variations ofa compound of formula (I) or a pharmaceutically acceptable salt thereof.An isotopic variation of a compound of formula (I), or apharmaceutically acceptable salt thereof, is defined as one in which atleast one atom is replaced by an atom having the same atomic number butan atomic mass different from the atomic mass usually found in nature.Examples of isotopes that can be incorporated into compounds of theinvention include isotopes of hydrogen, carbon, nitrogen, oxygen,fluorine and chlorine such as ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁷O, ¹⁸O, ¹⁸F and³⁶Cl, respectively. Certain isotopic variations of a compound of formula(I) or a salt or solvate thereof, for example, those in which aradioactive isotope such as ³H or ¹⁴C is incorporated, are useful indrug and/or substrate tissue distribution studies. Tritiated, i.e., ³H,and carbon-14, i.e., ¹⁴C, isotopes are particularly preferred for theirease of preparation and detectability. Further, substitution withisotopes such as deuterium, i.e., ²H, may afford certain therapeuticadvantages resulting from greater metabolic stability, for example,increased in vivo half-life or reduced dosage requirements and hence maybe preferred in some circumstances. Isotopic variations of a compound offormula (I), or a pharmaceutically salt thereof, can generally beprepared by conventional procedures such as by the illustrative methodsor by the preparations described in the Examples hereafter usingappropriate isotopic variations of suitable reagents.

The compounds of formula (I) and pharmaceutically acceptable saltsthereof may contain one or more asymmetric center (also referred to as achiral center) and may, therefore, exist as individual enantiomers,diastereomers, or other stereoisomeric forms, or as mixtures thereof.Chiral centers, such as chiral carbon atoms, may also be present in asubstituent such as an alkyl group. Where the stereochemistry of achiral center present in a compound of formula (I), or in any chemicalstructure illustrated herein, is not specified the structure is intendedto encompass all individual stereoisomers and all mixtures thereof.Thus, compounds of formula (I) and pharmaceutically acceptable saltsthereof containing one or more chiral center may be used as racemicmixtures, enantiomerically enriched mixtures, or as enantiomericallypure individual stereoisomers.

Individual stereoisomers of a compound of formula (I), or apharmaceutically acceptable salt thereof, which contain one or moreasymmetric center may be resolved by methods known to those skilled inthe art. For example, such resolution may be carried out (1) byformation of diastereoisomeric salts, complexes or other derivatives;(2) by selective reaction with a stereoisomer-specific reagent, forexample by enzymatic oxidation or reduction; or (3) by gas-liquid orliquid chromatography in a chiral environment, for example, on a chiralsupport such as silica with a bound chiral ligand or in the presence ofa chiral solvent. The skilled artisan will appreciate that where thedesired stereoisomer is converted into another chemical entity by one ofthe separation procedures described above, a further step is required toliberate the desired form. Alternatively, specific stereoisomers may besynthesized by asymmetric synthesis using optically active reagents,substrates, catalysts or solvents, or by converting one enantiomer tothe other by asymmetric transformation.

Certain compounds of the invention described herein possess an alphaamino acid ester that facilitates penetration of the compound throughthe cell wall. When inside the cell, the ester is hydrolysed byintracellular carboxyesterases to release the parent acid. Due to itscharge, the parent acid has reduced permeability and is thus retainedwithin the cell, where it becomes more concentrated leading to increasedpotency and duration of action. Even though compounds of the inventioncomprising an alpha amino acid ester are converted to theircorresponding carboxylic acid by intracellular esterases, both theesters and their corresponding acids function as as inhibitors of theBET family of bromodomain containing proteins. In one embodiment, acompound of the invention is capable of inhibiting the binding of one ormore of the four known BET family bromodomain containing proteins (e.g.BRD2, BRD3, BRD4 and BRDt) to, for example, an acetylated lysineresidue. In a further embodiment, the compound of formula (I), or apharmaceutically acceptable salt thereof, is capable of inhibiting thebinding of BRD4 to its cognate acetylated lysine residue. The compoundsof the invention may possess an improved profile over known BETinhibitors, for example, certain compounds may have one or more of thefollowing properties:

-   -   (i) potent BET inhibitory activity;    -   (ii) selectivity over other known bromodomain containing        proteins outside of the BET family of proteins;    -   (iii) selectivity for a particular BET family member over other        BET family members;    -   (iv) selectivity for one Binding Domain (i.e. BD1 over BD2 or        vice versa) for any given BET family member;    -   (v) improved developability (e.g. desirable solubility profile,        pharmacokinetics and pharmacodynamics); or    -   (vi) a reduced side-effect profile.

Further, certain compounds of the invention may inhibit other knownbromodomain containing proteins that are outside of the BET family ofproteins, such as, for example, bromodomain adjacent to zinc fingerdomain protein 2A (BAZ2A).

The compounds of the invention comprise a R₄ substituent attached ateither the 5- or 6-position of the benzimidazole core, which representsan alpha amino acid ester group and also captures the correspondingcarboxylic acids. The particular structure of the alpha amino acidesters of R₄ ensures that the ester is hydrolysed by cells containingcarboxyesterase hCE-1, and not by cells that contain othercarboxyesterases (such as hCE-2 and hCE-3) but not hCE-1. This propertyenables selective targeting of the compounds of the invention to cellsthat express hCE-1, such as macrophages, monocytes and dendritic cells.

Carboxyesterases hCE-2 and hCE-3 have a ubiquitous expression pattern,whereas hCE-1 is highly expressed in liver, lung and bone marrow but is,importantly, only found in certain types of cell, such as monocytes,macrophages and dendritic cells.

The structure of the alpha amino acid ester group, in particular thesubstitution pattern at positions R₇ and R₈, can determine the rate ofhydrolysis of the compound within cells that contain hCE-1, and thedesired rate of hydrolysis may differ depending on the selected route ofadministration.

hCE-1 is present in hepatocytes and, thus, for orally administeredcompounds, ester groups that have a slower rate of hydrolysis aredesirable to ensure that a sufficient amount of the compound enters thebloodstream after first pass metabolism.

In one embodiment of the present invention, a desirable rate ofhydrolysis for an orally administered compound may be obtained if R₇represents cycloalkyl, heterocycloalkyl, or —CR₁₃R₁₄R₁₅ wherein R₁₃ ishydrogen, hydroxyl, —CH₂OH, —CH₂C₁₋₃alkyl, halo, C₁₋₃alkyl, orC₁₋₃alkoxy wherein said C₁₋₃alkyl or C₁₋₃alkoxy may be optionallysubstituted with halo or hydroxyl; and R₁₄, and R₁₅ are independentlyhydrogen or C₁₋₃alkyl, with the proviso that at least two of R₁₃, R₁₄and R₁₅ are not hydrogen. In a further embodiment, R₇ representsisopropyl, —CH(CH₃)OH, sec-butyl, tert-butyl, tert-pentyl, sec-pentyl,3-pentyl or cycloalkyl. Similarly, R may represent —CHR₁₆R₁₇ wherein R₁₆is C₁₋₃alkyl and R₁₇ is C₁₋₆alkyl, cycloalkyl, heterocycloalkyl, whereinsaid C₁₋₆alkyl is optionally substituted with C₁₋₃alkoxy, or R₁₆ and R₁₇together with the carbon atom to which they attach form a cycloalkyl orheterocycloalkyl. In a further embodiment, R₈ represents isopropyl,sec-butyl, sec-pentyl, 3-pentyl, or cycloalkyl.

In one embodiment, the present invention is also includes eachcorresponding acid of the exemplified covalent conjugates that comprisean alpha amino acid ester (i.e. the covalent conjugates of Examples 1 to324 that comprises an alpha amino acid ester).

Statement of Use

Compounds of formula (I), or pharmaceutically acceptable salts thereof,are BET inhibitors and thus may have therapeutic utility in thetreatment of a variety of diseases or conditions related to systemic ortissue inflammation, inflammatory responses to infection or hypoxia,cellular activation and proliferation, lipid metabolism, fibrosis and inthe prevention and treatment of viral infections.

BET inhibitors may be useful in the treatment of a wide variety of acuteor chronic autoimmune or inflammatory conditions such as rheumatoidarthritis, osteoarthritis, acute gout, psoriasis, systemic lupuserythematosus, pulmonary arterial hypertension (PAH), multiplesclerosis, inflammatory bowel disease (Crohn's disease and Ulcerativecolitis), asthma, chronic obstructive airways disease, pneumonitis,myocarditis, pericarditis, myositis, eczema, dermatitis (includingatopic dermatitis), alopecia, vitiligo, bullous skin diseases,nephritis, vasculitis, hypercholesterolemia, atherosclerosis,Alzheimer's disease, depression, Sjögren's syndrome, sialoadenitis,central retinal vein occlusion, branched retinal vein occlusion,Irvine-Gass syndrome (post cataract and post-surgical), retinitispigmentosa, pars planitis, birdshot retinochoroidopathy, epiretinalmembrane, cystic macular edema, parafoveal telengiectasis, tractionalmaculopathies, vitreomacular traction syndromes, retinal detachment,neuroretinitis, idiopathic macular edema, retinitis, dry eye(keratoconjunctivitis Sicca), vernal keratoconjunctivitis, atopickeratoconjunctivitis, uveitis (such as anterior uveitis, pan uveitis,posterior uveitis, uveitis-associated macular edema), scleritis,diabetic retinopathy, diabetic macula edema, age-related maculardystrophy, hepatitis, pancreatitis, primary biliary cirrhosis,sclerosing cholangitis, Addison's disease, hypophysitis, thyroiditis,type I diabetes, giant cell arteritis, nephritis including lupusnephritis, vasculitis with organ involvement such as glomerulonephritis,vasculitis including giant cell arteritis, Wegener's granulomatosis,Polyarteritis nodosa, Behcet's disease, Kawasaki disease, Takayasu'sArteritis, pyoderma gangrenosum, vasculitis with organ involvement andacute rejection of transplanted organs. The use of BET inhibitors forthe treatment of rheumatoid arthritis is of particular interest.

In one embodiment, the acute or chronic autoimmune or inflammatorycondition is a disorder of lipid metabolism via the regulation of APO-A1such as hypercholesterolemia, atherosclerosis and Alzheimer's disease.

In another embodiment, the acute or chronic autoimmune or inflammatorycondition is a respiratory disorder such as asthma or chronicobstructive airways disease.

In another embodiment, the acute or chronic autoimmune or inflammatorycondition is a systemic inflammatory disorder such as rheumatoidarthritis, osteoarthritis, acute gout, psoriasis, systemic lupuserythematosus, multiple sclerosis or inflammatory bowel disease (Crohn'sdisease and ulcerative colitis).

In another embodiment, the acute or chronic autoimmune or inflammatorycondition is multiple sclerosis.

In a further embodiment, the acute or chronic autoimmune or inflammatorycondition is Type I diabetes.

BET inhibitors may be useful in the treatment of diseases or conditionswhich involve inflammatory responses to infections with bacteria,viruses, fungi, parasites or their toxins, such as sepsis, acute sepsis,sepsis syndrome, septic shock, endotoxaemia, systemic inflammatoryresponse syndrome (SIRS), multi-organ dysfunction syndrome, toxic shocksyndrome, acute lung injury, ARDS (adult respiratory distress syndrome),acute renal failure, fulminant hepatitis, burns, acute pancreatitis,post-surgical syndromes, sarcoidosis, Herxheimer reactions,encephalitis, myelitis, meningitis, malaria and SIRS associated withviral infections such as influenza, herpes zoster, herpes simplex andcoronavirus. In one embodiment, the disease or condition which involvesan inflammatory response to an infection with bacteria, a virus, fungi,a parasite or their toxins is acute sepsis.

BET inhibitors may be useful in the treatment of conditions associatedwith ischaemia-reperfusion injury such as myocardial infarction,cerebro-vascular ischaemia (stroke), acute coronary syndromes, renalreperfusion injury, organ transplantation, coronary artery bypassgrafting, cardio-pulmonary bypass procedures, pulmonary, renal, hepatic,gastro-intestinal or peripheral limb embolism.

BET inhibitors may be useful in the treatment of fibrotic conditionssuch as idiopathic pulmonary fibrosis, renal fibrosis, post-operativestricture, keloid scar formation, scleroderma (including morphea),cardiac fibrosis and cystic fibrosis.

BET inhibitors may be useful in the treatment of viral infections suchas herpes simplex infections and reactivations, cold sores, herpeszoster infections and reactivations, chickenpox, shingles, humanpapilloma virus (HPV), human immunodeficiency virus (HIV), cervicalneoplasia, adenovirus infections, including acute respiratory disease,poxyirus infections such as cowpox and smallpox and African swine fevervirus. In one embodiment, the viral infection is a HPV infection of skinor cervical epithelia. In another embodiment, the viral infection is alatent HIV infection.

BET inhibitors may be useful in the treatment of cancer, includinghematological (such as leukaemia, lymphoma and multiple myeloma),epithelial including lung, breast and colon carcinomas, midlinecarcinomas, mesenchymal, hepatic, renal and neurological tumours.

BET inhibitors may be useful in the treatment of one or more cancersselected from brain cancer (gliomas), glioblastomas, Bannayan-Zonanasyndrome, Cowden disease, Lhermitte-Duclos disease, breast cancer,inflammatory breast cancer, colorectal cancer, Wilm's tumor, Ewing'ssarcoma, rhabdomyosarcoma, ependymoma, medulloblastoma, colon cancer,head and neck cancer, kidney cancer, lung cancer, liver cancer,melanoma, squamous cell carcinoma, ovarian cancer, pancreatic cancer,prostate cancer, sarcoma cancer, osteosarcoma, giant cell tumor of bone,thyroid cancer, lymphoblastic T-cell leukemia, chronic myelogenousleukemia, chronic lymphocytic leukemia, hairy-cell leukemia, acutelymphoblastic leukemia, acute myelogenous leukemia, chronic neutrophilicleukemia, acute lymphoblastic T-cell leukemia, plasmacytoma,immunoblastic large cell leukemia, mantle cell leukemia, multiplemyeloma, megakaryoblastic leukemia, acute megakaryocytic leukemia,promyelocytic leukemia, mixed lineage leukaemia, erythroleukemia,malignant lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma,lymphoblastic T-cell lymphoma, Burkitt's lymphoma, follicular lymphoma,neuroblastoma, bladder cancer, urothelial cancer, vulval cancer,cervical cancer, endometrial cancer, renal cancer, mesothelioma,esophageal cancer, salivary gland cancer, hepatocellular cancer, gastriccancer, nasopharangeal cancer, buccal cancer, cancer of the mouth, GIST(gastrointestinal stromal tumor), NUT-midline carcinoma and testicularcancer.

In one embodiment, the cancer is a leukaemia, for example a leukaemiaselected from acute monocytic leukemia, acute myelogenous leukemia,chronic myelogenous leukemia, chronic lymphocytic leukemia and mixedlineage leukaemia (MLL). In another embodiment, the cancer isNUT-midline carcinoma. In another embodiment, the cancer is multiplemyeloma. In another embodiment, the cancer is a lung cancer such assmall cell lung cancer (SCLC). In another embodimnet, the cancer is aneuroblastoma. In another embodiment, the cancer is Burkitt's lymphoma.In another embodiment, the cancer is cervical cancer. In anotherembodiment, the cancer is esophageal cancer. In another embodiment, thecancer is ovarian cancer. In another embodiment, the cancer is breastcancer. In another embodiment, the cancer is colorectal cancer.

In one embodiment, the disease or condition for which a BET inhibitor isindicated is selected from diseases associated with systemicinflammatory response syndrome, such as sepsis, burns, pancreatitis,major trauma, haemorrhage and ischaemia. In this embodiment, the BETinhibitor would be administered at the point of diagnosis to reduce theincidence of SIRS, the onset of shock, multi-organ dysfunction syndrome,which includes the onset of acute lung injury, ARDS, acute renal,hepatic, cardiac or gastro-intestinal injury and mortality. In anotherembodiment, the BET inhibitor would be administered prior to surgical orother procedures associated with a high risk of sepsis, haemorrhage,extensive tissue damage, SIRS or MODS (multiple organ dysfunctionsyndrome). In a particular embodiment, the disease or condition forwhich a BET inhibitor is indicated is sepsis, sepsis syndrome, septicshock and endotoxaemia. In another embodiment, the BET inhibitor isindicated for the treatment of acute or chronic pancreatitis. In anotherembodiment, the BET inhibitor is indicated for the treatment of burns.

In a further aspect, the present invention also provides a compound offormula (I) or a pharmaceutically acceptable salt thereof for use intherapy.

In a further aspect, the present invention provides (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,of formula:

or a pharmaceutically acceptable salt thereof, for use in therapy.

In a further aspect, the present invention provides the1,2-ethanedisulphonic acid salt of (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,of formula:

for use in therapy.

In a further aspect, the present invention provides (2S,3R)-isopropyl2-(((1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,of formula:

or a pharmaceutically acceptable salt thereof, for use in therapy.

In a further aspect, the present invention provides a compound offormula (I) or a pharmaceutically acceptable salt thereof for use in thetreatment of diseases or conditions for which a bromodomain inhibitor,in particular a BET inhibitor, is indicated, including each and all ofthe above listed indications.

In a further aspect, the present invention also provides a compound offormula (I), or a pharmaceutically acceptable salt thereof, for use inthe treatment of autoimmune and inflammatory diseases, and cancer.

In a further aspect, the present invention provides a compound offormula (I), or a pharmaceutically acceptable salt thereof, for use inthe treatment of rheumatoid arthritis.

In a further aspect, the present invention is directed to a method oftreatment of an autoimmune or inflammatory disease or cancer, whichcomprises administering to a subject in need thereof, a therapeuticallyeffective amount of a compound of formula (I), or a pharmaceuticallyacceptable salt thereof.

In yet a further aspect, the present invention is directed to a methodof treating rheumatoid arthritis, which comprises administering to asubject in need thereof, a therapeutically effective amount of acompound of formula (I), or a pharmaceutically acceptable salt thereof.

In a further aspect, the present invention is directed to the use of acompound of formula (I), or a pharmaceutically acceptable salt thereof,in the manufacture of a medicament for use in the treatment of anautoimmune or inflammatory disease, or cancer.

In a further aspect, the present invention is directed to the use of acompound of formula (I), or a pharmaceutically acceptable salt thereof,in the manufacture of a medicament for use in the treatment ofrheumatoid arthritis.

Pharmaceutical Compositions/Routes of Administration/Dosages

While it is possible that for use in therapy, a compound of formula (I)as well as pharmaceutically acceptable salts thereof may be administeredas the raw chemical, it is common to present the active ingredient as apharmaceutical composition.

In a further aspect, there is provided a pharmaceutical compositioncomprising a compound of formula (I), or a pharmaceutically acceptablesalt thereof, and one or more pharmaceutically acceptable excipients. Ina further aspect, there is provided a pharmaceutical compositioncomprising a compound of formula (I), or a pharmaceutically acceptablesalt thereof, and a pharmaceutically acceptable excipient.

In a further aspect, there is provided a pharmaceutical compositioncomprising (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,of formula:

or a pharmaceutically acceptable salt thereof, and one or morepharmaceutically acceptable excipients.

In a further aspect, there is provided a pharmaceutical compositioncomprising the 1,2-ethanedisulphonic acid salt of (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,of formula:

In a further aspect, there is provided a pharmaceutical compositioncomprising (2S,3R)-Isopropyl2-(((1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,of formula:

or a pharmaceutically acceptable salt thereof, and one or morepharmaceutically acceptable excipients.

The excipient(s) must be pharmaceutically acceptable and be compatiblewith the other ingredients of the composition. In accordance withanother aspect of the invention there is also provided a process for thepreparation of a pharmaceutical composition including admixing acompound of formula (I), or a pharmaceutically acceptable salt thereof,with one or more pharmaceutically acceptable excipients. Thepharmaceutical composition can be used in the treatment of any of thediseases described herein.

Since the compounds of formula (I) are intended for use inpharmaceutical compositions it will be readily understood that they areeach preferably provided in substantially pure form, for example, atleast 85% pure, especially at least 98% pure (% in a weight for weightbasis).

Pharmaceutical compositions may be presented in unit dose formscontaining a predetermined amount of active ingredient per unit dose.Preferred unit dosage compositions are those containing a daily dose orsub-dose, or an appropriate fraction thereof, of an active ingredient.Such unit doses may therefore be administered more than once a day.

Pharmaceutical compositions may be adapted for administration by anyappropriate route, for example by the oral (including buccal orsublingual), rectal, inhaled, intranasal, topical (including buccal,sublingual or transdermal), ocular (including topical, intraocular,subconjunctival, episcleral, sub-Tenon), vaginal or parenteral(including subcutaneous, intramuscular, intravenous or intradermal)route. Such compositions may be prepared by any method known in the artof pharmacy, for example by bringing into association the activeingredient with the excipient(s).

In one aspect, the pharmaceutical composition is adapted for oraladministration.

In a further aspect, a compound of the invention may be formulated insuch a way as to facilitate delivery of said compound intracellularly.

Pharmaceutical compositions adapted for oral administration may bepresented as discrete units such as tablets or capsules; powders orgranules; solutions or suspensions in aqueous or non-aqueous liquids;edible foams or whips; or oil-in-water liquid emulsions or water-in-oilliquid emulsions.

Powders suitable for incorporating into tablets or capsules may beprepared by reducing the compound to a suitable fine size (e.g. bymicronisation) and mixing with a similarly prepared pharmaceuticalexcipient such as an edible carbohydrate, for example, starch ormannitol. Flavoring, preservative, dispersing and coloring agents, forexample, may also be present.

Capsules may be made by preparing a powder mixture, as described above,and filling formed gelatin sheaths. Glidants and lubricants such ascolloidal silica, talc, magnesium stearate, calcium stearate or solidpolyethylene glycol can be added to the powder mixture before thefilling operation. A disintegrating or solubilizing agent such asagar-agar, calcium carbonate or sodium carbonate can also be added toimprove the availability of the medicament when the capsule is ingested.

Moreover, when desired or necessary, suitable binders, glidants,lubricants, sweetening agents, flavours, disintegrating agents andcoloring agents can also be incorporated into the mixture. Suitablebinders include starch, gelatin, natural sugars such as glucose orbeta-lactose, corn sweeteners, natural and synthetic gums such asacacia, tragacanth or sodium alginate, carboxymethylcellulose,polyethylene glycol, waxes and the like. Lubricants used in these dosageforms include sodium oleate, sodium stearate, magnesium stearate, sodiumbenzoate, sodium acetate, sodium chloride and the like. Disintegratorsinclude starch, methyl cellulose, agar, bentonite, xanthan gum and thelike. Tablets are formulated, for example, by preparing a powdermixture, granulating or slugging, adding a lubricant and disintegrantand pressing into tablets. A powder mixture is prepared by mixing thecompound, suitably comminuted, with a diluent or base as describedabove, and optionally, with a binder such as carboxymethylcellulose, analiginate, gelatin, or polyvinyl pyrrolidone, a solution retardant suchas paraffin, a resorption accelerator such as a quaternary salt and/oran absorption agent such as bentonite, kaolin or dicalcium phosphate.The powder mixture can be granulated by wetting with a binder such assyrup, starch paste, acadia mucilage or solutions of cellulosic orpolymeric materials and forcing through a screen. As an alternative togranulating, the powder mixture can be run through the tablet machineand the result is imperfectly formed slugs broken into granules. Thegranules can be lubricated to prevent sticking to the tablet formingdies by means of the addition of stearic acid, a stearate salt, talc ormineral oil. The lubricated mixture is then compressed into tablets. Thecompounds of formula (I) and pharmaceutically acceptable salts thereofcan also be combined with a free flowing inert excipient and compressedinto tablets directly without going through the granulating or sluggingsteps. A clear or opaque protective coating consisting of a sealing coatof shellac, a coating of sugar or polymeric material and a polishcoating of wax can be provided. Dyestuffs can be added to these coatingsto distinguish different unit dosages.

Oral fluids such as solution, syrups and elixirs can be prepared indosage unit form so that a given quantity contains a predeterminedamount of the compound. Syrups can be prepared by dissolving thecompound in a suitably flavored aqueous solution, while elixirs areprepared through the use of a non-toxic alcoholic vehicle. Suspensionscan be formulated by dispersing the compound in a non-toxic vehicle.Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols andpolyoxy ethylene sorbitol ethers, preservatives, flavor additive such aspeppermint oil or natural sweeteners or saccharin or other artificialsweeteners, and the like can also be added. Compositions for oraladministration may be designed to provide a modified release profile soas to sustain or otherwise control the release of the therapeuticallyactive agent.

Where appropriate, dosage unit compositions for oral administration canbe microencapsulated. The composition may be prepared to prolong orsustain the release as for example by coating or embedding particulatematerial in polymers, wax or the like.

Pharmaceutical compositions for nasal or inhaled administration mayconveniently be formulated as aerosols, solutions, suspensions, gels ordry powders.

For pharmaceutical compositions suitable for and/or adapted for inhaledadministration, it is preferred that a compound of formula (I) or apharmaceutically acceptable salt thereof, is in a particle-size-reducedform e.g. obtained by micronisation. The preferable particle size of thesize-reduced (e.g. micronised) compound or salt is defined by a D50value of about 0.5 to about 10 microns (for example as measured usinglaser diffraction).

For pharmaceutical compositions suitable for and/or adapted for inhaledadministration, the pharmaceutical composition may be a dry powdercomposition or an aerosol formulation, comprising a solution or finesuspension of the active substance in a pharmaceutically acceptableaqueous or non-aqueous solvent. Dry powder compositions can comprise apowder base such as lactose, glucose, trehalose, mannitol or starch, thecompounds of formulae (I) or a pharmaceutically acceptable salt thereof(preferably in particle-size-reduced form, e.g. in micronised form), andoptionally a performance modifier such as L-leucine or another aminoacid and/or metal salt of stearic acid such as magnesium or calciumstearate. Preferably, the dry powder inhalable composition comprises adry powder blend of lactose e.g. lactose monohydrate and the compound offormula (I) or a salt thereof.

In one embodiment, a dry powder composition suitable for inhaledadministration may be incorporated into a plurality of sealed dosecontainers provided on medicament pack(s) mounted inside a suitableinhalation device. The containers may be rupturable, peelable orotherwise openable one-at-a-time and the doses of the dry powdercomposition administered by inhalation on a mouthpiece of the inhalationdevice, as known in the art. The medicament pack may take a number ofdifferent forms, for instance a disk-shape or an elongate strip.Representative inhalation devices are the DISKHALER™ inhaler device, theDISKUS™ inhalation device, and the ELLIPTA™ inhalation device, marketedby GlaxoSmithKline. The DISKUS™ inhalation device is, for example,described in GB 2242134A, and the ELLIPTA™ inhalation device is, forexample, described in WO 03/061743 A1 WO 2007/012871 A1 and/orWO2007/068896.

Pharmaceutical compositions adapted for parenteral administrationinclude aqueous and non-aqueous sterile injection solutions which maycontain anti-oxidants, buffers, bacteriostats and solutes which renderthe composition isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents. The compositions may be presented inunit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in a freeze-dried (lyophilized) conditionrequiring only the addition of the sterile liquid carrier, for examplewater for injections, immediately prior to use. Extemporaneous injectionsolutions and suspensions may be prepared from sterile powders, granulesand tablets.

Pharmaceutical compositions adapted for topical administration may beformulated as ointments, creams, suspensions, emulsions, lotions,powders, solutions, pastes, gels, foams, sprays, aerosols or oils. Suchpharmaceutical compositions may include conventional additives whichinclude, but are not limited to, preservatives, solvents to assist drugpenetration, co-solvents, emollients, propellants, viscosity modifyingagents (gelling agents), surfactants and carriers. In one embodimentthere is provided a pharmaceutical composition adapted for topicaladministration which comprises between 0.01-10%, or between 0.01-1% of acompound of formula (I)-(XVI), or a pharmaceutically acceptable saltthereof, by weight of the composition.

For treatments of the eye or other external tissues, for example mouthand skin, the compositions are preferably applied as a topical ointment,cream, gel, spray or foam. When formulated in an ointment, the activeingredient may be employed with either a paraffinic or a water-miscibleointment base. Alternatively, the active ingredient may be formulated ina cream with an oil-in-water cream base or a water-in-oil base.Pharmaceutical compositions adapted for topical administrations to theeye include eye drops wherein the active ingredient is dissolved orsuspended in a suitable carrier, especially an aqueous solvent.

A therapeutically effective amount of a compound of formula (I) or apharmaceutically acceptable salt thereof, will depend upon a number offactors including, for example, the age and weight of the subject, theprecise condition requiring treatment and its severity, the nature ofthe formulation, and the route of administration, and will ultimately beat the discretion of the attendant physician or veterinarian. In thepharmaceutical composition, each dosage unit for oral administrationpreferably contains from 0.01 to 1000 mg, more preferably 0.5 to 100 mg,of a compound of formula (I) or a pharmaceutically acceptable saltthereof, calculated as the free base.

The compounds of formula (I) and pharmaceutically acceptable saltsthereof may be employed alone or in combination with other therapeuticagents. Combination therapies according to the present invention thuscomprise the administration of at least one compound of formula (I) or apharmaceutically acceptable salt thereof, and the use of at least oneother therapeutically active agent. A compound of formula (I) orpharmaceutically acceptable salt thereof, and the other therapeuticallyactive agent(s) may be administered together in a single pharmaceuticalcomposition or separately and, when administered separately this mayoccur simultaneously or sequentially in any order.

In a further aspect, there is provided a combination product comprisinga compound of formula (I) or a pharmaceutically acceptable salt thereof,together with one or more other therapeutically active agents, andoptionally one or more pharmaceutically acceptable excipients.

It will be clear to a person skilled in the art that, where appropriate,the other therapeutic ingredient(s) may be used in the form of salts,for example as alkali metal or amine salts or as acid addition salts, oras solvates, for example hydrates, to optimise the activity and/orstability and/or physical characteristics, such as solubility, of thetherapeutic ingredient. It will be clear also that, where appropriate,the therapeutic ingredients may be used in optically pure form.

The combinations referred to above may conveniently be presented for usein the form of a pharmaceutical composition and thus pharmaceuticalcompositions comprising a combination as defined above together with apharmaceutically acceptable excipient.

General Synthetic Routes

The compounds of formula (I) and salts thereof may be prepared by themethodology described hereinafter, constituting further aspects of thisinvention.

Accordingly, there is provided a process for the preparation of acompound of formula (I), which process comprises the alkylation of acompound of formula (II):

Wherein R₂, R₃ and R₄ are as defined hereinbefore for a compound offormula (I). For example, a compound of formula (II) could be dissolvedin a solvent such as N,N-dimethylformamide, then treated with a suitablebase in the presence of an alkyl halide and heated at a suitabletemperature for an appropriate time to give, after purification,compounds of the formula (I) wherein R₁, R₂, R₃ and R₄ are as definedhereinbefore for a compound of formula (I).

There is provided a process for the preparation of a compound of formula(I), which process comprises cyclisation of a compound of formula (III):

Wherein R₃ and R₄ are as defined hereinbefore for a compound of formula(I). For example, a compound of formula (III) could be dissolved in asolvent mixture such as ethanol/water, then treated with a suitablealdehyde of formula (IV) in the presence of sodium dithionite and heatedat a suitable temperature for an appropriate time to give, afterpurification, compounds of the formula (I). The aldehydes mentionedabove are of general formula (IV) wherein R₁ and R₂ are as defined for acompound of formula (I).

There is provided a process for the preparation of a compound of formula(III), which process comprises the nucleophilic functionalisation of acompound of formula (V):

Wherein R₄ is as defined hereinbefore for a compound of formula (I). Forexample, a compound of formula (V) could be dissolved in a solvent suchas tetrahydrofuran then treated with a suitable amine containing R₃ asdefined hereinbefore for a compound of formula (I) in the presence of asuitable base such as triethylamine. The mixture would then be heated ata suitable temperature for an appropriate time to give, afterpurification, compounds of the formula (III).

There is provided a process for the preparation of a compound of formula(V), which process comprises the reductive amination of a compound offormula (VI):

Wherein (VI) is dissolved in a suitable solvent such as dichloromethaneto which is added an appropriately functionalised amine and an additivesuch as acetic acid. The mixture would be stirred at an appropriatetemperature for a specific time prior to the addition of a reducingagent such as sodium triacetoxyborohydride. The mixture would be stirredfor an appropriate time to give, after purification, compounds offormula (V) wherein R₄ is as defined hereinbefore for a compound offormula (I).

There is provided a process for the preparation of a compound of formula(VII), which process comprises the functionalisation of a compound offormula (VIII):

Wherein a compound of formula (VIII) could be dissolved in a solventsuch as dioxane, and then treated with a suitable amine containing R₃ asdefined hereinbefore for a compound of formula (I) in the presence of asuitable base such as triethylamine. The mixture would then be heated ata suitable temperature for an appropriate time to give, afterpurification, compounds of the formula (VII). The resulting compounds ofgeneral formula (VII) could then be reacted with aldehydes of formula(IV) in the presence of sodium dithionite in a suitable solvent mixturesuch as ethanol/water at a suitable temperature for an appropriate timeto give, after purification, compounds of the formula (IX).

The resulting compounds could then be further elaborated via sequentialalkylation, oxidation and reductive amination procedures by someoneskilled in the art to give further functionalised molecules fitting thegeneral formula (I) wherein R₁, R₂, R₃ and R₄ are as definedhereinbefore for a compound of formula (I).

There is provided a process for the preparation of a compound of formula(I), which process comprises the functionalisation of a compound offormula (X):

Wherein R₁, R₂ and R₃ are as defined hereinbefore for a compound offormula (I). For example, a compound of formula (X) could be dissolvedin a solvent mixture comprising acetonitrile and water, then treatedwith a suitable base in the presence of trimethylsulfonium iodide andheated at a suitable temperature for an appropriate time to give, afterworkup, functionalised intermediate compounds. These compounds couldthen be further elaborated by, for example, dissolution in a suitablesolvent such as tetrahydrofuran and addition of a Lewis acid such asboron trifluoride diethyl etherate. After stirring at an appropriatetemperature for an appropriate time, addition of a functionalised amine,a base and a reducing agent such as sodium triacetoxyborohydride wouldgive, after the appropriate reaction time, work up and purification,compounds of general formula (I) wherein R₁, R₂, R₃ and R₄ are asdefined hereinbefore for a compound of formula (I).

There is provided a process for the preparation of a compound of formula(X), which process comprises the functionalisation of a compound offormula (XI):

Wherein R₁, R₂ and R₃ are as defined hereinbefore for a compound offormula (I). For example, a compound of formula (XI) could be dissolvedin a solvent such as dichloromethane and treated with an appropriateoxidant for an appropriate time to give, after purification, compoundsof the formula (X).

There is provided a process for the preparation of a compound of formula(XII), which process comprises the functionalisation of a compound offormula (XIII):

Wherein a compound of formula (XIII) could be dissolved in a solventsuch as tetrahydrofuran, then treated with a suitable amine containingR₃ as defined hereinbefore for a compound of formula (I) in the presenceof a suitable base such as triethylamine. The mixture would then beheated at a suitable temperature for an appropriate time to give, afterpurification, compounds of the formula (XII). The resulting compounds ofgeneral formula (XII) could then be reacted with aldehydes of formula(IV) in the presence of sodium dithionite in a suitable solvent mixturesuch as ethanol/water at a suitable temperature for an appropriate timeto give, after purification, compounds of the formula (XI), wherein R₁,R₂ and R₃ are as defined hereinbefore for a compound of formula (I).

Further to this, there is provided a process for the preparation of acompound of formula (I), which process comprises the functionalisationof a compound of formula (X) wherein R₁, R₂ and R₃ are as definedhereinbefore for a compound of formula (I). For example, a compound offormula (X) could be dissolved in a solvent such as dichloromethanebefore being treated with an appropriately functionalised amine in thepresence of an additive such as triethylamine. After the appropriatereaction time, the resulting intermediate could be reduced followingaddition of a reducing agent such as sodium triacetoxyborohydride at anappropriate time and temperature, subsequent work up and purification ofthe mixture would give compounds of general formula (I) wherein R₁, R₂,R₃ and R₄ are as defined hereinbefore for a compound of formula (I).

Additionally, a process is provided for the preparation of compounds offormula (I) wherein R₁, R₂, R₃ and R₄ are as defined hereinbefore for acompound of formula (I) and R₄ contains a functionalised carboxylicacid. For example, a compound of general formula (I) could be dissolvedin a solvent mixture such as tetrahydrofuran/methanol/water then treatedwith a base such as lithium hydroxide for an appropriate time to give,after purification, a compound of general formula (I) wherein R₁, R₂, R₃and R₄ are as defined hereinbefore for a compound of formula (I).

Compounds IV, VI, VIII, and XIII are commercially available from, forexample, Sigma Aldrich.

EXAMPLES

Thus the following Examples serve to illustrate their preparation butare not to be considered as limiting the scope of the invention in anyway.

In the Intermediate and Example preparations, use of the phrase“prepared from: Intermediate X” or “prepared from Example Y” indicatewhere to find an example preparation for the compounds used (e.g.Intermediate X or Example Y), rather than the exact preparationnecessarily used.

Abbreviations

-   Ac Acetyl-   Bn Benzyl-   BOC tert-Butyloxycarbonyl-   dba Dibenzylideneacetone-   DCM Dichloromethane-   DIPEA N,N-diisopropylethylamine-   DMAP 4-Dimethylaminopyridine-   DMF N,N-dimethylformamide-   DMSO Dimethylsulfoxide-   EDC N-(3-Dimethylaminopropyl)-N-ethylcarbodiimide-   ee Enantiomeric excess-   Et Ethyl-   EtOAc Ethyl acetate-   EtOH Ethanol-   H or hr Hour(s)-   HATU O-(7-Azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium    hexafluorophosphate-   HOBt Hydroxybenzotriazole-   HPLC High-performance liquid chromatography-   IC₅₀ Half maximal inhibitory concentration-   IPA Isopropyl alcohol-   LCMS Liquid chromatography-mass spectrometry-   MDAP Mass-directed auto-preparative HPLC-   Me Methyl-   MPER Mammalian protein extraction reagent-   MS Mass spectrometry-   nBuLi n-Butyllithium-   NMP N-Methyl-2-pyrrolidone-   NMR Nuclear magnetic resonance-   Pd/C Palladium on carbon-   Pd(PPh₃)₄ Tetrakis(triphenylphosphine)palladium(0)-   Ph Phenyl-   ppm Parts per million-   pTSA para-Toluene sulfonic acid-   rt Retention time-   SCX Sulfonic acid, strong cation exchange-   SPE Solid phase extraction-   tBu Tertiary butyl-   TFA Trifluoroacetic acid-   t_(RET) Retention time-   THF Tetrahydrofuran-   UV Ultraviolet

Experimental Details NMR

¹H NMR spectra were recorded in either CDCl₃ or DMSO-d₆ on either aBruker DPX 400 or Bruker Avance DRX, Varian Unity 400 spectrometer orJEOL Delta all working at 400 MHz. The internal standard used was eithertetramethylsilane or the residual protonated solvent at 7.25 ppm forCDCl₃ or 2.50 ppm for DMSO-d₆.

LCMS System A

Column: 50 mm×2.1 mm ID, 1.7 μm Acquity UPLC BEH C₁₈Flow Rate: 1 mL/min.

Temp: 40° C.

UV detection range: 210 to 350 nmMass spectrum: Recorded on a mass spectrometer using alternative-scanpositive and negative mode electrospray ionisation

Solvents:

-   -   A: 0.1% v/v formic acid in water    -   B: 0.1% v/v formic acid acetonitrile

Gradient:

Time (min.) A % B % 0 97 3 1.5 0 100 1.9 0 100 2.0 97 3

System B

Column: 50 mm×2.1 mm ID, 1.7 μm Acquity UPLC BEH C₁₈Flow Rate: 1 mL/min.

Temp: 40° C.

UV detection range: 210 to 350 nmMass spectrum: Recorded on a mass spectrometer using alternative-scanpositive and negative mode electrospray ionisation

Solvents:

-   -   A: 10 mM ammonium bicarbonate in water adjusted to pH10 with        ammonia solution    -   B: acetonitrile

Gradient:

Time (min.) A % B % 0 99 1 1.5 3 97 1.9 3 97 2.0 0 100

System C

Column: 50 mm×2.1 mm ID, 1.7 μm Acquity UPLC CSH C₁₈Flow Rate: 1 mL/min.

Temp: 40° C.

UV detection range: 210 to 350 nmMass spectrum: Recorded on a mass spectrometer using alternative-scanpositive and negative mode electrospray ionisationThe solvents employed were:A=0.1% v/v solution of Trifluoroacetic Acid in Water.B=0.1% v/v solution of Trifluoroacetic Acid in Acetonitrile.

Gradient:

Time (min.) A % B % 0 95 5 1.5 5 95 1.9 5 95 2.0 95 5

System D

Column: 50 mm×2.1 mm ID, 1.7 μm Acquity UPLC CSH C₁₈Flow Rate: 1 mL/min.

Temp: 40° C.

UV detection range: 210 to 350 nmMass spectrum: Recorded on a mass spectrometer using alternative-scanpositive and negative mode electrospray ionisationThe solvents employed were:A=10 mM ammonium bicarbonate in water adjusted to pH10 with ammoniasolution.

B=Acetonitrile. Gradient:

Time (min.) A % B % 0 97 3 1.5 5 95 1.9 5 95 2.0 97 3

System E

Column: XBridge BEH C18 (50 mm×4.6 mm, 2.5 μm)Flow Rate: 1.3 mL/min.

Temp: 35° C. Solvents:

-   -   A: 5 mM Ammonium Bicarbonate in water (pH 10)    -   B: Acetonitrile

Gradient:

Time (min.) A % B % 0 95 5 0.5 95 5 1 85 15 3.3 2 98 6.0 2 98 6.1 95 56.5 95 5

System F

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 μm)Flow Rate: 0.6 mL/min.

Temp: 35° C. Solvents:

-   -   A: 0.1% Formic Acid in water    -   B: 0.1% Formic Acid in acetonitrile

Gradient:

Time (min.) A % B % 0 97 3 0.4 97 3 3.2 2 98 3.8 2 98 4.2 97 3 4.5 97 3

System G

Column: Acquity BEH C18 (50 mm×2.1 mm, 1.7 μm)Flow Rate: 0.45 mL/min.

Temp: 35° C. Solvents:

-   -   A: 0.05% Formic Acid in acetonitrile    -   B: 0.05% Formic Acid in water

Gradient:

Time (min.) A % B % 0 3 97 0.4 3 97 7.5 98 2 9.5 98 2 9.6 3 97

System H

Column: XBridge BEH C18 (50 mm×4.6 mm, 2.5 μm)Flow Rate: 1.3 mL/min.

Temp: 35° C. Solvents:

-   -   A: 5 mM Ammonium Bicarbonate in water (pH 10)    -   B: Acetonitrile

Gradient:

Time (min.) A % B % 0 95 5 0.5 95 5 1 85 15 6.0 2 98 9.0 2 98 9.5 95 510 95 5

System I

Column: 50 mm×2.1 mm ID, 1.7 μm Acquity UPLC CSH C₁₈Flow Rate: 1 mL/min.

Temp: 40° C.

UV detection range: 210 to 350 nmMass spectrum: Recorded on a mass spectrometer using alternative-scanpositive and negative mode electrospray ionisation

Solvents:

-   -   A: 0.1% v/v formic acid in water    -   B: 0.1% v/v formic acid acetonitrile

Gradient:

Time (min.) A % B % 0 97 3 1.5 5 95 1.9 5 95 2.0 97 3

System J

Column: 50 mm×2.1 mm ID, 1.7 μm Acquity UPLC CSH C₁₈Flow Rate: 1 mL/min.

Temp: 40° C.

UV detection range: 210 to 350 nmMass spectrum: Recorded on a mass spectrometer using alternative-scanpositive and negative mode electrospray ionisation

Solvents:

-   -   A: 10 mM ammonium bicarbonate in water adjusted to pH10 with        ammonia solution    -   B: acetonitrile

Gradient:

Time (min.) A % B % 0 97 3 0.05 97 3 1.5 5 95 1.9 5 95 2.0 97 3

Mass Directed Autopreparative HPLC (MDAP)

Mass directed autopreparative HPLC was undertaken under the conditionsgiven below. The UV detection was an averaged signal from wavelength of210 nm to 350 nm and mass spectra were recorded on a mass spectrometerusing alternate-scan positive and negative mode electrospray ionization.

Method A

Method A was conducted on a Sunfire C₁₈ column (typically 150 mm×30 mmi.d. 5 μm packing diameter) at ambient temperature. The solventsemployed were:

A=0.1% v/v solution of formic acid in waterB=0.1% v/v solution of formic acid in acetonitrile.

Method B

Method B was conducted on an XBridge C₁₈ column (typically 100 mm×30 mmi.d. 5 μm packing diameter) at ambient temperature. The solventsemployed were:

A=10 mM aqueous ammonium bicarbonate adjusted to pH 10 with ammoniasolution.B=acetonitrile.

Intermediate Preparation Intermediate 1:5-Methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde

5-Bromo-3-methylpyridin-2(1H)-one (commercially available from, forexample, Sigma Aldrich) (1.5 g, 7.98 mmol) was added to a flask whichwas purged with nitrogen. Anhydrous THF (75 mL) was added and thesolution was stirred under nitrogen, in dry-ice/acetone bath for 20 min.1.6 M n-Butyllithium in hexanes (14.96 mL, 23.93 mmol) was addeddropwise to the mixture and the reaction mixture was stirred undernitrogen in dry-ice/acetone bath for 3 hours. Anhydrous DMF (14.83 mL,191 mmol) was added dropwise and the reaction mixture was stirred undernitrogen in dry-ice/acetone bath for 1 hour. It was quenched usingsaturated aqueous ammonium chloride solution (30 mL) and allowed to warmto room temperature. The resulting slurry was partitioned between EtOAc(100 mL) and water (100 mL) and the layers were separated. The organiclayer was washed with brine (50 mL), dried, and evaporated under reducedpressure to give a pale yellow solid. The solid was triturated withdiethyl ether and the solid filtered was collected as the title compound(Batch 1, 210.8 mg). The aqueous layers from the previous extractionswere combined and re-extracted with DCM (4×75 mL and 4×100 mL). Thecombined organic layers were dried and evaporated under reduced pressureto give a pale yellow liquid. The liquid residue was azeotroped withtoluene (2×30 mL) and the solvents were removed under reduced pressureto give a yellow solid. The solid was triturated with diethyl ether, andthe solid filtered was collected as the title compound (Batch 2, 452.4mg). The total yield of the reaction was 61%. Batch 1: LCMS (System A):t_(RET)=0.41 min, MH⁺=138. Batch 2: LCMS (System A): t_(RET)=0.41 min,MH⁺=138.

Intermediate 2: 1,5-Dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde

A mixture of 5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (For anexample preparation see Intermediate 1, 5 g, 36.5 mmol) and potassiumcarbonate (10.08 g, 72.9 mmol) in DMF (50 mL) was cooled in an ice/waterbath, and methyl iodide (5.70 mL, 91 mmol) added dropwise. The reactionmixture was stirred for 15 min under nitrogen and then allowed to warmto room temperature and stirred for a further 2.5 hours. The solid wasremoved by filtration and the resulting solution evaporated underreduced pressure. The residue was partitioned between EtOAc (2×150 mL)and 1:1 water:saturated brine solution (150 mL). The organic layers werecombined, dried using a hydrophobic frit, and evaporated under reducedpressure. The sample was loaded in DCM and purified by silica gel columnchromatography (100 g column) using a gradient of 0-50% EtOAc incyclohexane. The appropriate fractions were combined and evaporatedunder reduced pressure to give the title compound (4.4 g, 80% yield) asan off-white solid. LCMS (System A): t_(RET)=0.46 min, MH⁺=152.

Intermediate 3: (S)-cyclopentyl 2-amino-4-methylpentanoate 4-methylbenzenesulfonate

A round bottom flask was charged with (S)-2-amino-4-methylpentanoic acid(5 g, 38.1 mmol), Cyclohexane (100 mL), tosic acid monohydrate (9.43 g,49.6 mmol) and cyclopentanol (35 mL, 386 mmol). A Dean-Stark condenserwas fitted and the mixture warmed to 130° C. to effect completedissolution. The mixture was stirred at this temperature over theweekend before being allowed to stand at room temperature for 7 days.The precipitated solid was isolated by filtration and washedsequentially with cyclohexane and ethyl acetate. The solid was dried invacuo to give the title compound (5.56 g) as a white solid. ¹H NMR (400MHz, METHANOL-d₄) δ 7.62-7.79 (m, 2H), 7.25 (d, J=7.83 Hz, 2H),5.15-5.42 (m, 1H), 3.97 (t, J=6.97 Hz, 1H), 2.39 (s, 3H), 1.42-2.10 (m,11H), 1.02 (d, J=7.09 Hz, 6H).

Intermediate 4: (2S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate

A mixture of (S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanoic acid(2.5 g, 11.51 mmol), diisopropylethylamine (2.97 g, 4.02 mL, 23.01mmol), 1-hydroxybenzotriazole hydrate (2.12 g, 13.84 mmol), EDC (2.65 g,13.81 mmol), and tetrahydrofuran-3-ol (10.14 g, 9.33 mL, 115 mmol) inDMF (20 mL) was stirred at room temperature overnight. The reactionmixture was partitioned between ethyl acetate (50 mL) and saturatedNaHCO₃ (50 mL). The organic phase was washed with 1M hydrochloric acid(50 mL), water (50 mL) and brine (50 mL). The organic phase was driedand evaporated to give the title compound (3.19 g) as a colourless oil.¹H NMR (400 MHz, CHLOROFORM-d) δ 5.34 (d, J=1.71 Hz, 1H), 4.93-5.09 (m,1H), 4.16-4.26 (m, 1H), 3.72-3.97 (m, 4H), 2.07-2.23 (m, 2H), 1.44 (s,9H), 0.82-1.02 (m, 6H).

Intermediate 5: (2S)-tetrahydrofuran-3-yl 2-amino-3-methylbutanoatehydrochloride

A solution of (2S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate (For a preparation seeIntermediate 4, 3.1 g, 10.79 mmol) in dioxan (5 mL) was treated with 4Mhydrogen chloride in dioxan (10 mL, 40 mmol). The reaction mixture wasstirred at room temperature for 24 hours. The solvent was evaporated andthe residue azeotroped with toluene (×3) to give the title compound(1.81 g), as a light brown oil. The crude product was used in subsequentreactions without purification.

Intermediate 6: (2S,3R)-2-hydroxy-2-methylpropyl2-((tert-butoxycarbonyl)amino)-3-hydroxybutanoate

In a microwave vial, 2,2-dimethyloxirane (2.058 mL, 23.17 mmol) wasadded to a mixture of(2S,3R)-2-((tert-butoxycarbonyl)amino)-3-hydroxybutanoic acid (1.016 g,4.63 mmol) and sodium bicarbonate (2.038 g, 24.26 mmol). The resultingmixture was heated under microwave irradiation at 120° C. for 30 min.Mixture was filtered on a bond eluent filter, using EtOAc as eluent.Volatiles were removed under reduced pressure to afford 1.75 g of aclear oil as a crude product. The crude product was purified by silicagel column chromatography (50 g Silica column), eluted with a 0-100%EtOAc in Cyclohexane gradient over 10 column volumes. The appropriatefractions were combined, and the volatiles were removed under reducedpressure to afford the title compound (0.8 g) as a clear oil. Theproduct was used crude in the next step without further purification.

Intermediate 7: (2S,3R)-2-hydroxy-2-methylpropyl2-amino-3-hydroxybutanoate

4.0M HCl in Dioxane (1 mL, 32.9 mmol) was added to(2S,3R)-2-hydroxy-2-methylpropyl2-((tert-butoxycarbonyl)amino)-3-hydroxybutanoate (For a preparation seeIntermediate 6, 85 mg, 0.292 mmol) and the resulting mixture was stirredat r.t. for 2.5 hours. The volatiles were removed under reduced pressureand the oil was triturated with Et₂O to give the title compound (55.8mg). Intermediate was used crude in following step without purification.

Intermediate 8: (2S,3R)-(S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-methoxybutanoate

(2S,3R)-2-((tert-butoxycarbonyl)amino)-3-methoxybutanoic acid (5000 mg,21.44 mmol), EDC (4931 mg, 25.7 mmol), DMAP (262 mg, 2.144 mmol) andHOBt (3939 mg, 25.7 mmol) was dissolved in DIPEA (7.49 mL, 42.9 mmol)and DMF (25 mL). The solution was stirred for 30 minutes prior to adding(S)-tetrahydrofuran-3-ol (17.39 mL, 215 mmol). The reaction mixture wasstirred overnight. The reaction mixture was partitioned between ethylacetate (150 mL) and saturated solution of sodium bicarbonate (150 mL).The organic fraction was isolated and the aqueous layer was re-extractedtwice with ethyl acetate (2×150 mL). The organic fractions werecombined, passed through a hydrophobic frit and concentrated underreduced pressure. The resultant oil was dissolved in DCM (3 mL) and thesolution was split into two which in turn were loaded onto silicacolumns (100 g columns). The product was eluted with a gradient of20-70% of ethyl acetate in cyclohexane. The appropriate fractions werecombined and dried in a vacuum oven to give the title compound (2156mg). ¹H NMR (400 MHz, DMSO-d₆) δ 5.84-5.99 (m, 1H), 5.25-5.33 (m, 1H),4.11 (dd, J=4.04, 8.59 Hz, 1H), 3.72-3.89 (m, 4H), 3.68 (s, 1H), 3.26(s, 3H), 2.12-2.26 (m, 1H), 1.86-2.00 (m, 1H), 1.38-1.46 (m, 9H),1.11-1.16 (m, 3H).

Intermediate 9: (2S,3R)-(S)-tetrahydrofuran-3-yl2-amino-3-methoxybutanoate hydrochloride

(2S,3R)-(S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-methoxybutanoate (For a preparation seeIntermediate 8, 5910 mg, 17.53 mmol) was dissolved in HCl (5M in IPA)(35.1 mL, 176 mmol) and the reaction mixture was stirred overnight. Thesolvent was removed under reduced pressure and the resulting oil wasplaced in a vacuum oven overnight to yield the title compound (3890 mg,93% yield) as an orange crystalline solid which was used in thesubsequent step without further purification.

Intermediate 10: (2S,3R)-Neopentyl 2-amino-3-hydroxybutanoate4-methylbenzenesulfonate

To a suspension of (2S,3R)-2-amino-3-hydroxybutanoic acid (2.27 g, 19.06mmol) and 4-methylbenzenesulfonic acid monohydrate (4.71 g, 24.77 mmol)in cyclohexane (100 mL) was added 2,2-dimethylpropan-1-ol (13.44 g, 152mmol). The reaction mixture was fitted with a Dean-Stark condenser andheated at 130° C. overnight. A white slurry was formed upon cooling toroom temperature, and it was evaporated under reduced pressure to removethe solvent. The white solid was dissolved in minimum amount of hotEtOAc and the clear solution was allowed to cool and then placed in anice/water bath. No crystallisation occurred. The solution was evaporatedunder reduced pressure and dried in a vacuum oven for four nights. Theresultant solid was recrystallised from EtOAc and allowed to cool,whereupon a white solid was formed. The resulting white solid wasremoved by filtration, washed with a little cold EtOAc, and dried in avacuum oven to give the title compound (6.0 g) as a white solid. ¹H NMR(d₆-DMSO, 293 K): δ 0.94 (s, 9H) 1.22 (d, J=6.3 Hz, 3H) 2.29 (s, 3H)3.84 (d, J=10.4 Hz, 1H) 3.92 (d, J=10.6 Hz, 1H) 3.99 (d, J=3.8 Hz, 1H)4.13-4.20 (m, 1H) 5.61 (d, J=4.3 Hz, 1H) 7.11 (m, J=7.8 Hz, 2H) 7.48 (m,J=8.1 Hz, 2H) 8.22 (br.s., 3H).

Intermediate 11: (2S,3R)-Isobutyl 2-amino-3-hydroxybutanoate4-methylbenzenesulfonate

To a suspension of (2S,3R)-2-amino-3-hydroxybutanoic acid (2.5 g, 20.99mmol) and 4-methylbenzenesulfonic acid monohydrate (5.19 g, 27.3 mmol)in cyclohexane (100 mL) was added 2-methylpropan-1-ol (15.50 mL, 168mmol). The reaction mixture was fitted with a Dean-Stark condenser andheated at 130° C. overnight. The reaction mixture was allowed to cool toroom temperature and the solvent removed under reduced pressure. Theresulting colourless liquid was dried in a vacuum oven for three nights.Initially, recrystallisation was attempted using EtOAc, but the sampleremained in solution and therefore the solvent was removed under reducedpressure. Trituration in diethyl ether was carried out twice, theresulting white solid was removed by filtration and dried in a vacuumoven to give the title compound (2.2 g) as a white solid. ¹H NMR(d₆-DMSO, 293 K): δ 0.92 (d, J=6.6 Hz, 6H) 1.21 (d, J=6.6 Hz, 3H)1.82-1.99 (m, 1H) 2.29 (s, 3H) 3.92-4.00 (m, 3H) 4.11-4.17 (m, 1H) 7.11(d, J=7.8 Hz, 2H) 7.48 (d, J=8.1 Hz, 2H) 8.23 (br.s., 3H).

Intermediate 12: (S)-isopropyl 2-amino-3-methylbutanoate4-methylbenzenesulfonate

(S)-2-amino-3-methylbutanoic acid (2.5 g, 21.34 mmol), tosic acid (5.28g, 27.7 mmol) and propan-2-ol (15 mL, 196 mmol) were dissolved incyclohexane (100 mL) and heated to 130° C. for 22 hrs. Solution wasallowed to cool to room temperature at which point a white solidprecipitated. This mixture was filtered under vacuum and the solidwashed several times with hexane. Solid was then placed in a vacuum ovenat 40° C. for 3 hrs. This solid was then added to cyclohexane (100 mL)along with tosic acid (1.76 g, 9.2 mmol) and propan-2-ol (3.94 g, 5.3mmol). Mixture was then heated to 130° C. for 20 hrs. Mixture wasallowed to cool to room temperature and filtered under gravity. Whitesolid was then placed in a vacuum oven at 40° C. for 24 hrs to give thetitle compound (5.98 g). ¹H NMR (400 MHz, METHANOL-d₄) δ 7.65-7.78 (m,2H), 7.25 (d, J=7.83 Hz, 2H), 5.10-5.19 (m, 1H), 3.87 (d, J=4.40 Hz,1H), 2.39 (s, 3H), 2.23-2.33 (m, 1H), 1.34 (d, J=6.11 Hz, 6H), 1.09 (dd,J=3.55, 6.97 Hz, 6H).

Intermediate 13: (S)-Cyclopentyl 2-aminobutanoate4-methylbenzenesulfonate

(S)-2-Aminobutanoic acid (3.0 g, 29.1 mmol), cyclopentanol (20.5 g, 238mmol) and 4-methylbenzenesulfonic acid monohydrate (7.19 g, 37.8 mmol)were added to cyclohexane (100 mL) and the reaction mixture was heatedto 125° C., whereupon complete dissolution was achieved. It was thenheated at this temperature for 24 hours. After cooling to roomtemperature, the volatile components were removed from the slurry underreduced pressure to give a white solid. The solid was recrystallisedfrom the minimum amount of hot EtOAc. The resulting crystals werefiltered and washed with a little cold EtOAc to give the title compound(6.30 g, 18.35 mmol, 63% yield) as a white solid. ¹H NMR (d₆-DMSO, 293K): δ 0.92 (t, J=7.5 Hz, 3H) 1.52-1.73 (m, 6H) 1.73-1.95 (m, 4H) 2.29(s, 3H) 3.90-4.01 (m, 1H) 5.18-5.22 (m, 1H) 7.11 (d, J=7.8 Hz, 2H) 7.48(d, J=8.1 Hz, 1H) 8.26 (br.s., 3H).

The following Intermediates were prepared in a similar manner toIntermediate 13 using the appropriate commercially available amino acidand alcohol starting materials:

Intermediate 14: (S)-tetrahydro-2H-pyran-4-yl 2-amino-4-methylpentanoate4-methylbenzenesulfonate

¹H NMR (d₆-DMSO, 293 K): δ 0.90 (d, J=2.9 Hz, 3H) 0.92 (d, J=2.9 Hz, 3H)1.49-1.78 (m, 5H) 1.84-1.93 (m, 1H) 2.29 (s, 3H) 3.46-3.54 (m, 2H)3.76-3.82 (m, 2H) 3.89-4.18 (m, 1H) 4.98-5.05 (m, 1H) 5.75 (s, 1H) 7.11(m, J=8.1 Hz, 2H) 7.48 (m, J=8.1 Hz, 2H) 8.29 (br.s., 3H), Yield: 2.8 g,68%

Intermediate 15: (S)-Neopentyl 2-amino-3-methylbutanoate4-methylbenzenesulfonate

¹H NMR (d₆-DMSO, 293 K): δ 0.94 (s, 9H), 0.97 (d, J=7.1 Hz, 3H), 1.01(d, J=6.9 Hz, 3H) 2.02-2.24 (m, 1H) 2.29 (s, 3H) 3.86 (d, J=10.3 Hz,1H), 3.90 (d, J=10.3 Hz, 1H) 3.98 (d, J=4.7 Hz, 1H) 7.11 (d, J=7.8 Hz,2H) 7.48 (d, J=8.1 Hz, 2H) 8.03-8.46 (br.s., 3H) Yield 3.9 g, 56%

Intermediate 16: (S)-Tetrahydro-2H-pyran-4-yl 2-amino-4-methylpentanoate4-methylbenzenesulfonate

¹H NMR (d₆-DMSO, 293 K): δ 0.90 (d, J=2.9 Hz, 3H) 0.92 (d, J=2.9 Hz, 3H)1.49-1.78 (m, 5H) 1.84-1.93 (m, 1H) 2.29 (s, 3H) 3.46-3.54 (m, 2H)3.76-3.82 (m, 2H) 3.89-4.18 (m, 1H) 4.98-5.05 (m, 1H) 5.75 (s, 1H) 7.11(m, J=8.1 Hz, 2H) 7.48 (m, J=8.1 Hz, 2H) 8.29 (br.s., 3H) Yield: 2.8 g,63%

Intermediate 17: (2S)-Tetrahydrofuran-3-yl 2-amino-4-methylpentanoate4-methylbenzenesulfonate

¹H NMR (d₆-DMSO, 293 K): δ 0.90 (d, J=2.2 Hz, 2H) 0.91 (d, J=2.2 Hz, 2H)1.54-1.77 (m, 3H) 1.86-2.00 (m, 1H) 2.13-2.25 (m, 1H) 2.29 (s, 3H)3.69-3.87 (m, 4H) 3.95-4.02 (m, 1H) 5.33-5.39 (m, 1H) 7.11 (d, J=8.1 Hz,2H) 7.47 (d, J=8.1 Hz, 2H) 8.28 (br.s., 3H) Yield: 3.37 g, 59%

Intermediate 18: (2S)-1-Methoxypropan-2-yl 2-amino-4-methylpentanoate4-methylbenzenesulfonate (Mixture of Diastereomers)

¹H NMR (d₆-DMSO, 293 K): δ 0.85-1.01 (m, 6H) 1.19 (d, J=4.7 Hz, 1.3H)1.21 (d, J=4.9 Hz, 1.8H) 1.50-1.68 (m, 2H) 1.68-1.85 (m, 1H) 2.29 (s,3H) 3.25 (s, 1.7H) 3.27 (s, 1.2H) 3.36-3.51 (m, 2H) 3.93-4.02 (m, 1H)5.03-5.17 (m, 1H) 7.11 (d, J=7.8 Hz, 2H) 7.48 (d, J=8.1 Hz, 2H) 8.29(br.s., 3H) Yield: 950 mg, 15%

Intermediate 19: (S)-Neopentyl 2-amino-4-methylpentanoate4-methylbenzenesulfonate

¹H NMR (d₆-DMSO, 293 K): δ 0.90 (d, J=3.7 Hz, 2H) 0.91-0.94 (m, 12H)1.54-1.81 (m, 3H) 2.29 (s, 3H) 3.84 (d, J=10.5 Hz, 1H) 3.89 (d, J=10.5Hz, 1H) 3.98-4.08 (m, 1H) 7.11 (d, J=7.8 Hz, 2H) 7.48 (d, J=8.1 Hz, 2H)8.20-8.41 (br.s., 3H) Yield: 4.91 g, 86%

Intermediate 20: (S)-Cyclobutyl 2-amino-4-methylpentanoate4-methylbenzenesulfonate

¹H NMR (d₆-DMSO, 293 K): δ 0.90 (d, J=2.0 Hz, 3H) 0.91 (d, J=2.0 Hz, 3H)1.54-1.84 (m, 5H) 1.96-2.15 (m, 2H) 2.23-2.39 (m, 5H) 3.97 (t, J=7.1 Hz,1H) 4.98-5.06 (m, 1H) 7.11 (m, J=7.8 Hz, 2H) 7.48 (m, J=8.1 Hz, 2H) 8.27(br.s., 3H) Yield: 5.7 g, 92%

Intermediate 21: (S)-Isopropyl 2-amino-4-methylpentanoate4-methylbenzenesulfonate

¹H NMR (d₆-DMSO, 293 K): δ 0.89 (d, J=2.0 Hz, 3H) 0.91 (d, J=2.3 Hz, 3H)1.22-1.26 (m, 6H) 1.54-1.66 (m, 2H) 1.67-1.81 (m, 1H) 2.29 (s, 3H) 3.93(t, J=7.1 Hz, 1H) 4.91-5.10 (m, 1H) 7.12 (m, J=8.1 Hz, 2H) 7.48 (m,J=8.1 Hz, 2H) 8.27 (br.s., 3H) Yield 4.2 g, 70%

Intermediate 22: (S)-Cyclopentyl2-amino-2-(tetrahydro-2H-pyran-4-yl)acetate 4-methylbenzenesulfonate

¹H NMR (d₆-DMSO, 293 K): δ 1.19-1.36 (m, 1H) 1.40-1.53 (m, 2H) 1.53-1.74(m, 7H) 1.82-1.92 (m, 2H) 1.97-2.11 (m, 1H) 2.29 (s, 3H) 3.20-3.31 (m,2H) 3.81-3.96 (m, 3H) 5.19-5.24 (s, 1H) 7.11 (d, J=7.8 Hz, 2H) 7.48 (d,J=8.1 Hz, 2H) 8.30 (br.s., 3H) Yield: 1.77 g, 35%

Intermediate 23: (S)-Cyclopentyl 2-amino-4-methoxybutanoate4-methylbenzenesulfonate

¹H NMR (d₆-DMSO, 293 K): δ 1.49-1.62 (m, 2H) 1.62-1.77 (m, 4H) 1.76-1.92(m, 2H) 2.00 (q, J=6.1 Hz, 2H) 2.29 (s, 3H) 3.22 (s, 3H) 3.40-3.43 (m,partially obscured by solvent peak) 3.92-4.09 (m, 1H) 5.16-5.20 (m, 1H)7.11 (d, J=7.8 Hz, 2H) 7.48 (d, J=8.1 Hz, 2H) 8.24 (br.s., 3H) Yield:4.70 g, 56%

Intermediate 24: (S)-Cyclopentyl 2-amino-3-methylbutanoate4-methylbenzenesulfonate

(S)-2-Amino-3-methylbutanoic acid (800 mg, 6.83 mmol), cyclopentanol(5.07 ml, 55.9 mmol) and 4-methylbenzenesulfonic acid monohydrate (1.689g, 8.88 mmol) were suspended in cyclohexane (15 mL) and the reactionmixture was heated to 95° C. with a Dean-Stark condenser attached.Complete dissolution was achieved, and the reaction mixture was heatedat 95° C. overnight. No water had been collected in the Dean-Starkapparatus. The temperature of the reaction mixture was raised to 110°C., and the reaction mixture was heated at 110° C. overnight. Thereaction mixture was cooled to room temperature to give a white slurry.The solvent was removed under reduced pressure to give a white solid.This crude material was dissolved in a minimum volume of hot EtOAc andthe solution was allowed to cool. The resulting white solid wasfiltered, washed with a small amount of EtOAc, and dried to give thetitle compound (1.89 g, 5.28 mmol, 77% yield) as an off-white solid. ¹HNMR (d₆-DMSO, 293 K): δ 0.94 (d, J=7.1 Hz, 3H) 0.96 (d, J=7.1 Hz, 3H)1.54-1.76 (m, 6H) 1.81-1.95 (m, 2H) 2.05-2.20 (m, 1H) 2.29 (s, 3H)3.85-3.90 (m, 1H) 5.20-5.24 (m, 1H) 7.11 (d, J=7.8 Hz, 2H) 7.47 (d,J=8.1 Hz, 2H) 8.23 (br.s., 3H).

Intermediate 25: (2S,3R)-Isobutyl 2-amino-3-hydroxybutanoate4-methylbenzenesulfonate

To a suspension of (2S,3R)-2-amino-3-hydroxybutanoic acid (2.5 g, 20.99mmol) and 4-methylbenzenesulfonic acid monohydrate (5.19 g, 27.3 mmol)in cyclohexane (100 mL) was added 2-methylpropan-1-ol (15.50 ml, 168mmol). The reaction mixture was fitted with a Dean-Stark condenser andheated at 130° C. overnight. The reaction mixture was allowed to cool toroom temperature and the solvent removed under reduced pressure. Theresulting colourless liquid was dried in a vacuum oven for three nights.Initially, recrystallisation was attempted using EtOAc, but the sampleremained in solution and therefore the solvent was removed under reducedpressure. Trituration in diethyl ether was again unsuccessful, so thesample (with ether still present) was stood overnight open to the air,with a pasteur pipette and spatula left in the gum to attempt to inducecrystallisation. A small amount of solid had formed around the spatulaand pipette, so trituration in diethyl ether was again attempted, andwas this time successful. The resulting white solid was removed byfiltration and dried in a vacuum oven to give the title compound (2.2 g,6.33 mmol, 30% yield) as a white solid. ¹H NMR (d₆-DMSO, 293 K): δ 0.92(d, J=6.6 Hz, 6H) 1.21 (d, J=6.6 Hz, 3H) 1.82-1.99 (m, 1H) 2.29 (s, 3H)3.92-4.00 (m, 3H) 4.11-4.17 (m, 1H) 7.11 (d, J=7.8 Hz, 2H) 7.48 (d,J=8.1 Hz, 2H) 8.23 (br.s., 3H).

Intermediate 26: Cyclopentyl 1-aminocyclobutanecarboxylate4-methylbenzenesulfonate

1-Aminocyclobutanecarboxylic acid (1.66 g, 14.42 mmol),4-methylbenzenesulfonic acid monohydrate (3.57 g, 18.74 mmol) andcyclopentanol (10.16 g, 118 mmol) were added to cyclohexane (100 mL) andthe reaction mixture was heated to 130° C. Full dissolution wasachieved, and the reaction mixture was heated at this temperature for 3days. The solvent had evaporated due to the condenser not being fittedproperly. The resulting pale brown solid was recrystallised from EtOAc,filtered, washed, and dried to give the tosic acid salt of the startingmaterial amino acid. This recovered starting material (3.7726 g, 13.13mmol), cyclopentanol (10.16 g, 118 mmol) and 4-methylbenzenesulfonicacid monohydrate (0.823 g, 4.33 mmol) were added to cyclohexane (100 mL)and the reaction mixture was heated at 130° C. for 5 days. The resultingslurry was evaporated under reduced pressure and dried in a vacuum oven.The solid was recrystallised from hot EtOAc, filtered, washed, and driedto give the title compound (3.31 g, 9.31 mmol, 65% yield) as a whitesolid. ¹H NMR (d₆-DMSO, 293 K): δ 1.55-1.65 (m, 2H) 1.65-1.77 (m, 4H)1.81-1.93 (m, 2H) 1.97-2.05 (m, 2H) 2.29 (s, 3H) 2.30-2.39 (m, 2H)2.39-2.49 (m, 2H) 5.21-5.25 (m, 1H) 7.12 (d, J=8.3 Hz, 2H) 7.48 (d,J=8.1 Hz, 2H) 8.50 (br.s., 3H).

Intermediate 27:(2S,3S)-2-(((benzyloxy)carbonyl)amino)-3-hydroxybutanoic acid

To a stirred solution of (2S,3S)-2-amino-3-hydroxybutanoic acid (1 g,8.39 mmol) and potassium carbonate (2.90 g, 20.99 mmol) in water (18 mL)was added at 0° C. benzyl carbonochloridate (1.438 mL, 10.07 mmol). Thereaction mixture was stirred at room temperature for 21 h. The reactionmixture was washed by Et₂O and the aqueous acidified to pH 0 by additionof 2M aq. HCl. The resulting milky white solution was extracted 3 timesby EtOAc, concentrated in vacuo to give a colourless oil. Toluene wasadded to the oil and concentrated in vacuo to give the title compound(1.80 g) as a colourless oil. LCMS (System A): t_(RET)=0.68 min,MH⁺=254.

Intermediate 28: (2S,3S)-(S)-tetrahydrofuran-3-yl2-(((benzyloxy)carbonyl)amino)-3-hydroxybutanoate

To a stirred solution of(2S,3S)-2-(((benzyloxy)carbonyl)amino)-3-hydroxybutanoic acid (for apreparation see Intermediate 27, 0.920 g, 3.45 mmol) in DMF (5 mL) wasadded (S)-tetrahydrofuran-3-ol (2.343 mL, 34.5 mmol), EDC (0.794 g, 4.14mmol), DIPEA (1.205 mL, 6.90 mmol), DMAP (0.417 g, 3.42 mmol) and1-hydroxybenzotriazole hydrate (0.634 g, 4.14 mmol). The resultingsolution was stirred overnight and the reaction mixture was partitionedbetween EtOAc and aq. sat. NaHCO₃. The organic phase was washed with 2MHCl, passed through a hydrophobic frit, and concentrated in vacuo togive a colourless oil. The oil was dissolved in DCM and purified bysilica gel chromatography eluting with Cyclohexane:EtOAc (15-75%). Theappropriate fractions were combined and evaporated in vacuo to give thetitle compound as a colourless oil. The total yield of the reaction was46%. LCMS (System A): t_(RET)=0.79 min, MH⁺=324.

Intermediate 29: (2S,3S)-(S)-tetrahydrofuran-3-yl2-amino-3-hydroxybutanoate Hydrochloride

To a vacuum degassed black suspension of(2S,3S)-(S)-tetrahydrofuran-3-yl2-(((benzyloxy)carbonyl)amino)-3-hydroxybutanoate (for a preparation seeIntermediate 28 0.508 g, 1.571 mmol) and 10% Pd/C (0.167 g, 0.157 mmol)in Ethanol (20 mL) was added a hydrogen atmosphere. The reaction mixturewas stirred for 5h 30 min, passed through a celite cartridge,concentrated in vacuo and dried under a stream of nitrogen to obtain abrown oil. Et2O (2 mL) and 1M HCl in Et2O (1.46 mL) were added to thebrown oil, concentrated under a stream of nitrogen, and dried in vacuoto give the title compound as a brown solid. Compound used at thispurity.

Intermediate 30: (S)-(S)-Tetrahydrofuran-3-yl 2-amino-4-methylpentanoate

Tosic acid monohydrate (7.73 g, 40.6 mmol) was added to a stirredsuspension of L-leucine (4.1 g, 31.3 mmol) and(S)-(+)-3-Hydroxytetrahydrofuran (10 mL, 147 mmol) in cyclohexane (105mL). The resulting suspension was heated 110 OC with Dean-Starkapparatus for 2 days and cooled to room temperature with stirring. Theresulting solid was slurried in additional cyclohexane (100 mL),filtered and dried in vacuo to a brown solid (18 g). The solid wasslurried in toluene (100 mL) and heated to 60° C. The resulting solutionwas cooled to room temperature, filtered and the combined filtrates fromthe toluene and cyclohexane washes were combined with the filteredsolid. The resulting suspension was washed (3×aqueous saturated sodiumbicarbonate, 1×brine), dried over MgSO4, and evaporated in vacuo to abrown oil. The oil was loaded on to an SPE (silica, 100 g) and elutedwith 0-5% (2 M ammonia in methanol) in DCM. The clean, productcontaining fractions were evaporated in vacuo to give the title compound(2.58 g) as a light brown oil. LCMS (System C): t_(RET)=0.49 min,MH⁺=202

Intermediate 31: (2S,3R)-Isopropyl 2-amino-3-hydroxybutanoate4-methylbenzenesulfonate

To a suspension of (2S,3R)-2-amino-3-hydroxybutanoic acid (commerciallyavailable from, for example, Sigma Aldrich) (10 g, 84 mmol) incyclohexane (100 mL), 2-propanol (51.7 mL, 672 mmol) and4-methylbenzenesulfonic acid hydrate (20.76 g, 109 mmol) was added atroom temperature. A Dean-Stark apparatus was fitted and the reactionmixture was stirred at 105° C. for 4 days. The reaction mixture wasevaporated in vacuo to give a colourless oil (53 g). The oil wasrecrystallized from EtOAc (20 mL) to obtain a white solid (49 g). Thesolid was dried in a vacuum oven overnight to obtain the title compound(31.9 g) as a waxy white solid. Intermediate is ˜85% pure, used at thispurity level in subsequent reactions. ¹H NMR (d₆-DMSO): δ 1.20 (d, J=6.6Hz, 3H), 1.25 (dd, J=3.2, 6.4 Hz, 6H), 2.29 (s, 3H), 3.82-3.92 (m, 1H),4.05-4.16 (m, 2H), 5.02 (m, 1H), 7.11 (d, J=7.9 Hz, 2H), 7.42-7.51 (m,2H), 8.20 (br. s., 3H).

Intermediate 31a: (2S,3R)-isopropyl 2-amino-3-hydroxybutanoatehydrochloride

(2S,3R)-isopropyl 2-amino-3-hydroxybutanoate 4-methylbenzenesulfonicacid salt (for an example preparation see Intermediate 31) (90 g) wasdissolved in water (200 mL) and basified with saturated sodium carbonatesolution (200 mL), then extracted with dichloromethane (5×200 mL) andthe organic layer dried and evaporated in vacuo to give a pale yellowoil. The product was dissolved in dichloromethane and 4.0M hydrogenchloride in dioxan (100 mL) was added, then the mixture was stirred for1 hour, then evaporated in vacuo and dried in a vacuum oven overnight togive (2S,3R)-isopropyl 2-amino-3-hydroxybutanoate hydrochloride (33.5 g,169 mmol) as a colourless crystalline solid. ¹H NMR (400 MHz, DMSO-d₆) δppm 8.44 (br s, 3H), 5.65 (d, J=10 Hz, 1H), 4.95-5.04 (m, 1H), 4.04-4.13(m, 1H), 3.80 (d, J=10 Hz, 1H), 1.18-1.29 (m, 9H).

Intermediate 32: (2S,3R)-Cyclobutyl 2-amino-3-hydroxybutanoate4-methylbenzenesulfonate

(2S,3R)-2-amino-3-hydroxybutanoic acid (15 g, 126 mmol), cyclobutanol(72.6 g, 1007 mmol) and p-toluenesulfonic acid monohydrate (31.1 g, 164mmol) were combined in cyclohexane (480 mL) and heated to 140° C. forfour nights with Dean-Stark apparatus attached. In total 8.4 mL waterwas removed. The reaction was concentrated in vacuo. The residue wastriturated with Et₂O (115 mL) for 2 hours. The resulting solid wascollected by filtration and washed with Et₂O (˜50 mL) then dried in avacuum oven to give the title compound (40.0064 g, 110 mmol) as a whitesolid. ¹H NMR (d₆-DMSO): δ 1.20 (d, J=6.6 Hz, 3H), 1.54-1.71 (m, 1H),1.71-1.88 (m, 1H), 1.98-2.16 (m, 2H), 2.23-2.38 (m, 5H), 3.92 (d, J=3.9Hz, 1H), 4.06-4.21 (m, 1H), 4.98-5.06 (m, 1H), 5.63 (d, J=4.4 Hz, 1H),7.11 (d, J=7.8 Hz, 2H), 7.41-7.52 (m, 2H), 8.21 (br. s., 2H).

Intermediate 33: (2S,3R)-(S)-sec-Butyl2-((tert-butoxycarbonyl)amino)-3-hydroxybutanoate

To a solution of(2S,3R)-2-((tert-butoxycarbonyl)amino)-3-hydroxybutanoic acid (2.95 g,13.46 mmol) in DMF (10 mL) was added HOBT (2.473 g, 16.15 mmol), EDC(3.10 g, 16.15 mmol), DMAP (0.164 g, 1.346 mmol) and DIPEA (4.70 mL,26.9 mmol). The reaction mixture was stirred at room temperature for 4nights. The reaction mixture was concentrated under reduced pressure,and the residue partitioned between EtOAc (2×150 mL) and saturatedaqueous sodium bicarbonate solution (150 mL). The organic layers werecombined, washed with 2 M aqueous hydrochloric acid (150 mL) and brine(150 mL), dried using a hydrophobic frit, and evaporated under reducedpressure. The sample was loaded in DCM and purified by SPE (silica, 100g) using a gradient of 0-100% EtOAc in cyclohexane. The appropriatefractions were combined and evaporated under reduced pressure to givethe title compound (2.0 g, 7.26 mmol) as a colourless gum. LCMS (SystemB): t_(RET)=1.01 min, MH⁺=276

Intermediate 34: (2S,3R)-(S)-sec-Butyl 2-amino-3-hydroxybutanoatehydrochloride

To a solution of (2S,3R)-(S)-sec-butyl2-((tert-butoxycarbonyl)amino)-3-hydroxybutanoate (For an examplepreparation see Intermediate 33, 2.0 g, 7.26 mmol) in 1,4-dioxane (16mL) was added 4 M hydrochloric acid in 1,4-dioxane (4.41 mL, 145 mmol)and the reaction mixture stirred at room temperature overnight. Thereaction mixture was evaporated under reduced pressure. The material wasdissolved in 1,4-dioxane (4 mL) and 4 M hydrochloric acid in 1,4-dioxane(4.41 mL, 145 mmol) was added. The reaction mixture was stirred at roomtemperature for 3 nights. The reaction mixture was evaporated underreduced pressure to leave the title compound (1.5 g, 7.09 mmol) as alight brown gum. ¹H NMR (d₆-DMSO): 0.89 (t, J=7.5 Hz, 3H), 1.22 (s, 3H),1.23 (s, 3H), 1.53-1.65 (m, 2H), 3.87 (d, J=3.8 Hz, 1H), 4.10-4.17 (m,1H), 4.81-4.91 (m, 1H), 5.50-5.73 (br.s., 1H), 8.36 (br.s., 3H).

Intermediate 35: (2S)-tetrahydrofuran-3-yl 2-aminobutanoatehydrochloride

A solution of (2S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)butanoate (for a preparation seeintermediate 36, 3.0 g, 10.98 mmol) in dioxan (5 mL) was treated with 4Mhydrogen chloride in dioxan (10 mL, 40 mmol). The reaction mixture wasstirred at room temperature for 24 hours. The solvent was evaporated andthe residue azeotroped with toluene (×3) to give the title compound(1.87 g, 8.92 mmol, 81% yield), as a colourless solid. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 8.55 (br s, 3H), 5.32-5.38 (m, 1H), 3.92-3.96 (m, 1H),3.70-3.84 (m, 4H). 2.13-2.24 (m, 2H), 1.88-2.00 (m, 1H), 1.78-1.87 (m,1H), 0.92 (t, J=12.0 Hz, 3H).

Intermediate 36: (2S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)butanoate

A mixture of (S)-2-((tert-butoxycarbonyl)amino)butanoic acid (2.5 g,12.3 mmol), diisopropylethylamine (3.18 g, 4.3 mL, 24.6 mmol),1-hydroxybenzotriazole hydrate (2.26 g, 14.76 mmol), EDC (2.83 g, 14.76mmol), and tetrahydrofuran-3-ol (10.84 g, 9.97 mL, 123 mmol) in DMF (20mL) was stirred at room temperature overnight. The reaction mixture waspartitioned between ethyl acetate (50 mL) and saturated NaHCO₃ (50 mL).The organic phase was washed with 1M hydrochloric acid (50 mL), water(50 mL) and brine (50 mL). The organic phase was dried and evaporated togive the title compound (3.07 g, 11.23 mmol, 91% yield), as a colourlessoil. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 5.30-5.34 (m, 1H), 5.00-5.04 (m,1H), 4.16-4.22 (m, 1H), 3.92-3.94 (m, 1H), 2.12-2.22 (m, 1H). 1.93-2.03(m, 1H), 1.78-1.84 (m, 1H), 1.60-1.69 (m, 1H), 1.43 (s, 9H), 0.92 (t,J=12.0 Hz, 3H).

Intermediate 37: (2S)-1-methoxypropan-2-yl 2-aminobutanoatehydrochloride

A solution of (2S)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)butanoate (for a preparation seeintermediate 38, 2.98 g, 10.82 mmol) in dioxan (5 mL) was treated with4M hydrogen chloride in dioxan (10 mL, 40 mmol). The reaction mixturewas stirred at room temperature for 24 hours. The solvent was evaporatedand the residue azeotroped with toluene (×3) to give(2S)-1-methoxypropan-2-yl 2-aminobutanoate hydrochloride (2.13 g, 10.06mmol, 93% yield) as a light brown oil. ¹H NMR (400 MHz, DMSO-d₆) δ ppm8.64 (br s, 3H), 5.04-5.13 (m, 1H), 3.89-3.93 (m, 1H), 3.56 (s, 3H),3.25 (d, J=10.0 Hz, 2H) 1.79-1.90 (m, 2H), 1.18 (dd, J=12 Hz, 4 Hz, 3H),0.93 (dt, J=10.0 Hz, 4 Hz, 3H).

Intermediate 38: (2S)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)butanoate

A mixture of (S)-2-((tert-butoxycarbonyl)amino)butanoic acid (2.5 g,12.3 mmol), diisopropylethylamine (3.18 g, 4.3 mL, 24.6 mmol),1-hydroxybenzotriazole hydrate (HOBt) (2.26 g, 14.76 mmol), EDC (2.83 g,14.76 mmol), and 1-methoxy-2-propanol (11.09 g, 12.02 mL, 123 mmol) inDMF (20 mL) was stirred at room temperature overnight. The reactionmixture was partitioned between ethyl acetate (50 mL) and saturatedNaHCO₃ (50 mL). The organic phase was washed with 1M hydrochloric acid(50 mL), water (50 mL) and brine (50 mL). The organic phase was driedand evaporated to give the title compound (2.98 g, 10.82 mmol, 88%yield), as a colourless oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 5.05-5.15 (m,2H), 4.20-4.30 (m, 1H), 3.37-3.48 (m, 2H), 3.34 (d, J=10.0 Hz, 1H),1.80-1.90 (m, 1H). 1.64-1.72 (m, 1H), 1.78-1.84 (m, 1H), 1.44 (s, 9H),1.22-1.27 (m, 3H), 0.89-0.96 (m, 3H).

Intermediate 39: (2S,3R)-(S)-Tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-hydroxybutanoate

To a solution of(2S,3R)-2-((tert-butoxycarbonyl)amino)-3-hydroxybutanoic acid (5 g, 22.8mmol) in N,N-dimethylformamide (22.5 mL) was added diisopropylethylamine(7.97 mL, 45.6 mmol), 1-hydroxybenzotriazole hydrate (4.19 g, 27.4mmol), EDC (5.25 g, 27.4 mmol), DMAP (0.28 g, 2.26 mmol) and(S)-tetrahydrofuran-3-ol (18.3 mL, 228 mmol). The reaction mixture wasstirred at RT for 72 h. The reaction mixture was partitioned betweenethyl acetate (200 mL) and saturated aqueous sodium hydrogen carbonate(200 mL). The organic phase was separated and washed with 2M aqueous HCl(160 mL), water (160 mL) and brine (160 mL). The organic phase was driedand evaporated to give the crude product as a clear oil. The oil wasdissolved in dichloromethane (2 mL) and loaded onto 2×100 g SNAP silicacolumn. The crude material on silica was purified by Biotage SP4 using agradient of 10-50% ethyl acetate in cyclohexane over 24 CV. Fractionscontaining pure product were collected and the solvent removed underreduced pressure and further dried under high vacuum overnight to yield(2S,3R)-(S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-hydroxybutanoate (2.81 g, 9.42 mmol,41% yield) as a clear gum. ¹H NMR δ (500 MHz, CDCl₃) ppm: 5.38 (1H, t,J=5.4 Hz), 5.29 (1H, d, J=7.7 Hz), 4.36-4.27 (1H, m), 4.23 (1H, d, J=8.5Hz), 3.98-3.81 (4H, m), 2.25-2.13 (1H, m), 2.10-2.01 (1H, m), 1.64 (1H,br. s), 1.46 (9H, s), 1.26 (3H, d, J=6.6 Hz).

Intermediate 40: (2S,3R)-(S)-tetrahydrofuran-3-yl2-amino-3-hydroxybutanoate, Hydrochloride

To a solution of (2S,3R)-(S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-hydroxybutanoate (for a preparation seeIntermediate 39, 570 mg, 1.97 mmol) in 1,4-dioxane (5.5 mL) at roomtemperature was added hydrochloric acid (4 M in dioxane) (1.97 mL, 7.88mmol). The reaction mixture was stirred at room temperature for 16 h.The reaction (monitored by TLC) showed conversion, but not tocompletion. An additional portion of hydrochloric acid (4 M in dioxane)(1.97 mL, 7.88 mmol) was added and stirred for 6 h. Hydrochloric acid (4M in dioxane) (1.97 mL, 7.88 mmol) was again added, stirred for 2 h andleft to stand overnight. The solvent was removed in vacuo and theproduct further dried in the vacuum oven to yield(2S,3R)-(S)-tetrahydrofuran-3-yl 2-amino-3-hydroxybutanoate,hydrochloride (470 mg, 1.979 mmol, 100% yield) as a yellow gum. ¹H NMR δ(400 MHz, DMSO-d6) ppm: 8.46 (3H, br. s.), 5.66 (1H, br. s.), 5.35 (1H,td, J=4.2, 2.3 Hz), 4.17-4.08 (1H, m), 3.89 (1H, d, J=3.8 Hz), 3.85-3.69(4H, m), 2.24-2.13 (1H, m), 2.00-1.91 (1H, m), 1.22 (3H, d, J=6.6 Hz).

Intermediate 41: (2S)-1-methoxypropan-2-yl 2-amino-3-methylbutanoatehydrochloride

A solution of (2S)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate (for a preparation seeIntermediate 42, 3.13 g, 10.82 mmol) in dioxan (5 mL) was treated with4M hydrogen chloride in dioxan (10 mL, 40 mmol). The reaction mixturewas stirred at room temperature for 24 hours. The solvent was evaporatedand the residue azeotroped with toluene (×3) to give the title compound(1.84 g, 8.15 mmol, 75% yield) as a light brown oil. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 8.64 (br s, 3H), 5.05-5.15 (m, 1H), 3.78-3.81 (m, 1H),3.33-3.43 (m, 2H), 3.23-3.25 (m, 3H), 2.15-2.24 (m, 1H), 1.19 (dd, J=12Hz, 4 Hz, 3H), 0.98-1.02 (m, 3H), 0.94 (dd, J=10.0 Hz, 3 Hz, 3H).

Intermediate 42: ((2S)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate

A mixture of (S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanoic acid(2.5 g, 11.51 mmol), diisopropylethylamine (2.97 g, 4.02 mL, 23.01mmol), 1-hydroxybenzotriazole hydrate (2.12 g, 13.84 mmol), EDC (2.65 g,13.81 mmol), and 1-methoxypropan-2-ol (10.37 g, 11.25 mL, 115 mmol) inDMF (20 mL) was stirred at room temperature overnight. The reactionmixture was partitioned between ethyl acetate (50 mL) and saturatedNaHCO₃ (50 mL). The organic phase was washed with 1M hydrochloric acid(50 mL), water (50 mL) and brine (50 mL). The organic phase was driedand evaporated to give the title compound (3.13 g, 10.82 mmol, 94%yield), as a colourless oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 5.00-5.15 (m,2H), 4.16-4.22 (m, 1H), 3.34-3.47 (m, 2H), 3.30-3.33 (m, 3H), 1.41 (s,9H), 1.21 (d, J=12 Hz, 3H), 0.94 (d, J=12 Hz, 3H), 0.84-0.88 (m, 3H).

Intermediate 43: (2S)-1-methoxypropan-2-yl 2-amino-3,3-dimethylbutanoatehydrochloride 4M Hydrogen chloride in dioxan (10 mL, 40 mmol) was addedto (2S)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoate (for a preparationsee Intermediate 44, 2.6 g, 8.63 mmol). The reaction mixture was stirredat room temperature for 24 hours. The solvent was evaporated and theresidue azeotroped with toluene (×3) to give the title compound (1.83 g,7.63 mmol, 89% yield) as a pale yellow oil. ¹H NMR (400 MHz, DMSO-d₆) δppm 8.55 (br s, 3H), 5.05-5.16 (m, 1H), 3.32-3.45 (m, 3H), 3.23-3.25 (m,3H), 1.19 (m, 3H), 1.02 (s, 9H).

Intermediate 44: (2S)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoate

A mixture of (S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoicacid (2.5 g, 10.8 mmol), diisopropylethylamine (2.79 g, 3.78 mL, 21.6mmol), 1-hydroxybenzotriazole hydrate (1.99 g, 12.97 mmol), EDC (2.49 g,12.97 mmol), and 1-methoxypropan-2-ol (9.74 g, 10.57 mL, 108 mmol) inDMF (20 mL) was stirred at room temperature overnight. The reactionmixture was partitioned between ethyl acetate (50 mL) and saturatedNaHCO₃ (50 mL). The organic phase was washed with 1M hydrochloric acid(50 mL), water (50 mL) and brine (50 mL). The organic phase was driedand evaporated to give the title compound (2.65 g, 8.73 mmol, 81% yield)as a colourless oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 5.08-5.18 (m, 2H),4.04-4.11 (m, 1H), 3.32-3.48 (m, 5H), 1.43 (s, 9H), 1.23 (d, J=12 Hz,3H), 0.97 (s, 9H).

Intermediate 45: (2S)-tetrahydrofuran-3-yl 2-amino-3,3-dimethylbutanoatehydrochloride

4M hydrogen chloride in dioxan (10 mL, 40 mmol) was added to(2S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoate (for a preparationsee Intermediate 46, 2.6 g, 8.63 mmol). The reaction mixture was stirredat room temperature for 24 hours. The solvent was evaporated and theresidue azeotroped with toluene (×3) to give the title compound ((1.78g, 7.49 mmol, 87% yield) as a colourless hygroscopic solid. ¹H NMR (400MHz, DMSO-d₆) δ ppm 8.58 (br s, 3H), 5.33-5.37 (m, 1H), 3.65-3.84 (m,5H), 2.12-2.22 (m, 1H), 1.94-2.03 (m, 1H), 1.01 (s, 9H).

Intermediate 46: (2S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoate

A mixture of (S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoicacid (2.5 g, 10.8 mmol), diisopropylethylamine (2.79 g, 3.78 mL, 21.6mmol), 1-hydroxybenzotriazole hydrate (1.99 g, 12.97 mmol), EDC (2.49 g,12.97 mmol), and tetrahydrofuran-3-ol (9.52 g, 8.76 mL, 123 mmol) in DMF(20 mL) was stirred at room temperature overnight. The reaction mixturewas partitioned between ethyl acetate (50 mL) and saturated NaHCO₃ (50mL). The organic phase was washed with 1M hydrochloric acid (50 mL),water (50 mL) and brine (50 mL). The organic phase was dried andevaporated to give the title compound (2.7 g, 8.96 mmol, 83% yield) as acolourless oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 5.30-5.35 (m, 1H),5.05-5.12 (m, 1H), 3.79-4.09 (m, 4H), 1.97-2.22 (m, 2H), 1.43 (s, 9H),0.97 (s, 9H).

Intermediate 47: (2S)-tetrahydrofuran-3-yl2-amino-3-hydroxy-3-methylbutanoate hydrochloride

A solution of (2S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-methylbutanoate (for apreparation see Intermediate 48, 320 mg, 1.05 mmol) in dioxan (2 mL) wastreated with 4M hydrogen chloride in dioxan (2 mL, 8 mmol). The reactionmixture was stirred at room temperature for 24 hours. The solvent wasevaporated. The residue was triturated with diethyl ether, a gum formed.The supernatant was decanted off and the residual solvent evaporated togive the title compound (250 mg, 1.043 mmol, 99% yield) as a colourlessfoam. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.41 (br s, 3H), 5.49-5.56 (m,1H), 5.32-5.38 (m, 1H), 3.70-3.85 (m, 5H), 2.11-2.23 (m, 1H), 1.93-2.02(m, 1H), 1.29-1.34 (m, 3H), 1.16-1.19 (m, 3H).

Intermediate 48: (2S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-methylbutanoate

A solution of(S)-2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-methylbutanoic acid (250mg, 1.072 mmol) in tetrahydrofuran (5 mL) was treated withtriphenylphosphine (309 mg, 1.18 mmol) and diisopropyl azodicarboxylate(238 mg, 229 μL, 1.18 mmol). The mixture was stirred at room temperaturefor 5 minutes then tetrahydrofuran-3-ol (472 mg, 434 μL, 5.36 mmol) wasadded. The reaction mixture was stirred at room temperature for 18hours. The solvent was evaporated. The residue was dissolved in ethylacetate (15 mL). The solution was washed with water (2×10 mL) and brine(10 mL). The organic phase was dried and evaporated. The residue waschromatographed [40% ethyl acetate/hexane] to give the title compound(142 mg, 0.468 mmol, 43.7% yield) as a colourless oil. ¹H NMR (400 MHz,CDCl₃) δ ppm 5.33-5.44 (m, 2H), 4.14-4.20 (m, 1H), 3.83-3.97 (m, 4H),2.13-2.25 (m, 1H), 2.00-2.12 (m, 1H), 1.44 (s, 9H), 1.26-1.29 (m, 6H).

Intermediate 49: (S)-(S)-tetrahydrofuran-3-yl 2-amino-3-methylbutanoatehydrochloride

A solution of (S)-(S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate (for a preparation seeIntermediate 50, 2.73 g, 9.50 mmol) in ethyl acetate (5 mL) was treatedwith 4M hydrogen chloride in dioxan (5 mL). The reaction mixture wasstirred at room temperature for 24 hours. The solvent was evaporated.Attempted trituration with diethyl ether did not give a solid. Thesolvent was evaporated to give the title compound (1.98 g, 8.85 mmol,93% yield) as a colourless oil. Sample solidified on standing at roomtemperature for several days. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.69 (brs, 3H), 5.32-5.37 (m, 1H), 3.70-3.84 (m, 5H), 2.12-2.24 (m, 2H),1.93-2.02 (m, 1H), 0.99 (t, J=12 Hz, 3H), 0.94 (t, J=12 Hz, 3H).

Intermediate 50: (S)-(S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate

A mixture of (S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanoic acid(2.5 g, 11.51 mmol), diisopropylethylamine (2.97 g, 4.02 mL, 23.01mmol), 1-hydroxybenzotriazole hydrate (HOBt) (2.12 g, 13.84 mmol), EDC(2.65 g, 13.81 mmol), and (S)-tetrahydrofuran-3-ol (5.07 g, 3.91 mL,57.5 mmol) in DMF (20 mL) was stirred at room temperature overnight. Thereaction mixture was partitioned between ethyl acetate (50 mL) andsaturated NaHCO₃ (50 mL). The organic phase was washed with 1Mhydrochloric acid (50 mL), water (50 mL) and brine (50 mL). The organicphase was dried and evaporated to give the title compound (2.73 g, 9.50mmol, 83% yield), as a colourless oil. ¹H NMR (400 MHz, CDCl₃) δ ppm5.30-5.35 (m, 1H), 4.90-5.04 (m, 1H), 4.16-4.20 (m, 1H), 3.75-3.93 (m,4H), 2.08-2.22 (m, 2H), 1.98-2.07 (m, 1H), 1.43 (s, 9H), 0.95 (t, J=12Hz, 3H), 0.88 (t, J=12 Hz, 3H).

Intermediate 51: (S)-cyclopentyl 2-amino-3-hydroxy-3-methylbutanoatehydrochloride

A solution of (S)-cyclopentyl2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-methyl butanoate (for apreparation see Intermediate 52, 330 mg, 1.09 mmol) in ethyl acetate (2mL) was treated with 4M HCl in dioxan (2 mL). The reaction mixture wasstirred at room temperature overnight. The solvent was evaporated.Attempted trituration with hexane/diethyl ether did not give pureproduct. The compound was dissolved in methanol and loaded onto an SCXcolumn. The column was washed with methanol then eluted with 2M ammoniain methanol. The NH₃/methanol fractions were evaporated. The residue wasdissolved in diethyl ether (5 mL) and the solution was treated with 1Mhydrogen chloride in diethyl ether (0.5 mL). The solvent was evaporatedto give the title compound (258 mg, 1.085 mmol, 99% yield) as ahygroscopic colourless gum. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.33 (br s,3H), 5.50-5.54 (m, 1H), 5.17-5.23 (m, 1H), 3.68-3.77 (m, 1H), 3.30-3.37(m, 1H), 1.78-1.89 (m, 2H), 1.53-1.74 (m, 5H), 1.17 (s, 6H).

Intermediate 52: (S)-cyclopentyl2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-methylbutanoate

A solution of(S)-2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-methylbutanoic acid (500mg, 2.14 mmol) in tetrahydrofuran (5 mL) was treated withtriphenylphosphine (618 mg, 2.36 mmol) and diisopropylazodicarboxylate(477 mg, 458 μL, 2.36 mmol). The reaction mixture was stirred at roomtemperature for 10 minutes then cyclopentanol (923 mg, 973 μL, 10.72mmol) was added. The reaction mixture was stirred at room temperaturefor 24 hours. The solvent was evaporated. The residue was dissolved inethyl acetate (10 mL). The solution was washed with saturated NaHCO₃solution (10 mL), 1M hydrochloric acid (10 mL), water (10 mL) and brine(10 mL). The organic phase was dried and evaporated. The residue waschromatographed [30% ethyl acetate/hexane] to give the title compound(330 mg, 1.095 mmol, 51.1% yield) as a colourless gummy solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 6.48 (br s, 1H), 5.20-5.43 (m, 1H), 4.93-5.01 (m,1H), 4.05-4.36 (m, 1H), 1.50-1.88 (m, 6H), 1.45 (s, 6H), 1.23-1.27 (m,9H).

Intermediate 53: (2S)-1-methoxypropan-2-yl2-amino-3-hydroxy-3-methylbutanoate hydrochloride

A solution of (2S)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-methylbutanoate (for apreparation see Intermediate 54, 480 mg, 1.57 mmol) in ethyl acetate (2mL) was treated with 4M HCl in dioxan (2 mL). The reaction mixture wasstirred at room temperature overnight. The solvent was evaporated. Thecompound was dissolved in methanol and loaded onto an SCX column. Thecolumn was washed with methanol then eluted with 2M ammonia in methanol.The NH₃/methanol fractions were evaporated. The residue was dissolved indiethyl ether (5 mL) and the solution was treated with 1M hydrogenchloride in diethyl ether (0.5 mL). The solvent was evaporated to givethe title compound (315 mg, 1.303 mmol, 83% yield) as a hygroscopiccolourless gum. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.27 (br s, 3H),5.48-5.54 (m, 1H), 5.06-5.16 (m, 1H), 3.74-3.82 (m, 1H), 3.38-3.47 (m,1H), 3.27 and 3.25 (2×s, 3H. OMe) 1.36 and 1.32 (2×s, 3H), 1.20 (d, J=8Hz, 3H), 1.17 and 1.14 (2×s, 3H).

Intermediate 54: (2S)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-methylbutanoate

A solution of(S)-2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-methylbutanoic acid (500mg, 2.14 mmol) in tetrahydrofuran (5 mL) was treated withtriphenylphosphine (618 mg, 2.36 mmol) and diisopropylazodicarboxylate(477 mg, 458 μL, 2.36 mmol). The reaction mixture was stirred roomtemperature for 10 minutes then 1-methoxypropan-2-ol (966 mg, 1.05 mL,10.72 mmol) was added. The reaction mixture was stirred at roomtemperature for 24 hours. The solvent was evaporated. The residue wasdissolved in ethyl acetate (10 mL). The solution was washed withsaturated NaHCO₃ solution (10 mL), 1M hydrochloric acid (10 mL), water(10 mL) and brine (10 mL). The organic phase was dried and evaporated.The residue was chromatographed [40% ethyl acetate/hexane]. Hexane (10mL) was added to the product and stirred for 30 minutes. The solid wasfiltered off, washed with hexane and dried to give the title compound(487 mg, 1.595 mmol, 74.4% yield), as a colourless solid. ¹H NMR (400MHz, CDCl₃) δ ppm 5.16-5.43 (m, 1H), 4.10-4.30 (m, 1H), 2.38-2.50 (m,2H), 3.35 and 3.37 (2×s, 3H), 1.60 (s, 3H), 1.45 (s, 3H), 1.25-1.29 (m,12H).

Intermediate 55: (S)-cyclopentyl 2-amino-2-cyclopropylacetate

20 v/v piperidine in DMF (1 mL) was added to a solution of(S)-cyclopentyl2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-cyclopropylacetate (fora preparation see Intermediate 56, 240 mg, 0.59 mmol) in DMF (1 mL). Thereaction mixture was stirred at room temperature for 18 hours. Thereaction mixture was partitioned between ethyl acetate (15 mL) and water(15 mL). The organic phase was separated, washed with water (2×10 mL),dried and evaporated. The residue was dissolved in methanol and loadedon to an SCX column. The column was washed with methanol (3CV) and theneluted with 2M ammonia in methanol (5CV). The ammonia/MeOH fraction wasevaporated. The residue was dissolved in dichloromethane (2 mL). 1.0MHydrogen chloride in diethyl ether (1 mL) was added. The solvent wasevaporated and the residue triturated with diethyl ether to give asolid. The solid was dissolved in ethyl acetate (20 mL) and the solutionwashed with saturated NaHCO₃ and brine. The organic phase was dried andevaporated. The residue was chromatographed [0-4%methanol/dichloromethane] to give the title compound (43 mg, 0.235 mmol,39.6% yield) as a colourless oil. ¹H NMR (400 MHz, CDCl₃) δ ppm5.18-5.25 (m, 1H), 2.78-2.82 (m, 1H), 1.80-1.93 (m, 2H), 1.56-1.78 (m,6H), 0.93-1.03 (m, 1H), 0.41-0.58 (m, 2H), 0.27-0.34 (m, 1H).

Intermediate 56: (S)-cyclopentyl2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-cyclopropylacetate

Dicyclohexylcarbodiimide (147 mg, 0.711 mmol) was added to a stirredsolution of(S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-cyclopropylaceticacid (200 mg, 0.593 mmol) in cyclopentanol (2 mL, large excess). Thereaction mixture was stirred at room temperature for 24 hours. Thereaction mixture was partitioned between ethyl acetate (20 mL) and water(20 mL). The organic phase was separated, washed with water and brine.Dried and evaporated. The residue was chromatographed [10-30% ethylacetate/hexane] to give the title compound (246 mg, 0.607 mmol, 102%yield) as a colourless solid. LCMS (System A): t_(RET)=1.42 min; MH⁺406.

Intermediate 57: (S)-cyclopentyl 2-amino-3-cyclopropylpropanoate

20 v/v piperidine in DMF (1 mL) was added to (S)-cyclopentyl2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-cyclopropylpropanoate(for a preparation see Intermediate 58, 270 mg, 0.59 mmol). The reactionmixture was stirred at room temperature for 18 hours. The reactionmixture was partitioned between ethyl acetate (15 mL) and saturatedNaHCO₃ solution (15 mL). The organic phase was separated, washed withwater (2×10 mL), and brine (10 mL). The organic phase was dried andevaporated. The residue was chromatographed [0-4%methanol/dichloromethane] to give the title compound (48 mg, 0.243 mmol,37.8% yield) as a colourless oil. ¹H NMR (400 MHz, CDCl₃) δ ppm5.11-5.18 (m, 1H), 3.42-3.46 (m, 1H), 3.24-3.28 (m, 1H), 1.76-1.86 (m,1H), 1.45-1.77 (m, 8H), 0.65-0.75 (m, 1H), 0.38-0.46 (m, 2H), 0.04-0.08(m, 2H).

Intermediate 58: (S)-cyclopentyl2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-cyclopropylpropanoate

Cyclopentanol (2 mL, large excess) was added to(S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-cyclopropylpropanoicacid (250 mg, 0.711 mmol). The mixture was stirred and treated withN-Ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) (211 mg, 0.854mmol). The reaction mixture was stirred at room temperature for 24hours. The reaction mixture was partitioned between ethyl acetate (10mL) and saturated NaHCO₃ solution (10 mL). The organic phase wasseparated, washed with water and brine. Dried and evaporated. Theresidue was chromatographed [10-30% ethyl acetate/hexane] to give thetitle compound (270 mg, 0.644 mmol, 90% yield).

LCMS (System A): t_(RET)=1.45 min; MH⁺ 420.

Intermediate 59: (R)-cyclopentyl 2-amino-3-cyclopropylpropanoate

20 v/v piperidine in DMF (1 mL) was added to (R)-cyclopentyl2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-cyclopropylpropanoate(for a preparation see Intermediate 60, 258 mg, 0.615 mmol). Thereaction mixture was stirred at room temperature for 18 hours. Thereaction mixture was partitioned between ethyl acetate (15 mL) andsaturated NaHCO₃ solution (15 mL). The organic phase was separated,washed with water (2×10 mL), and brine (10 mL). The organic phase wasdried and evaporated. The residue was chromatographed [0-4%methanol/dichloromethane] to give the title compound (59 mg, 0.299 mmol,48.6% yield) as a colourless oil. ¹H NMR (400 MHz, CDCl₃) δ ppm5.11-5.18 (m, 1H), 3.42-3.46 (m, 1H), 3.24-3.28 (m, 1H), 1.76-1.86 (m,1H), 1.45-1.77 (m, 8H), 0.65-0.75 (m, 1H), 0.38-0.46 (m, 2H), 0.04-0.08(m, 2H).

Intermediate 60: (R)-cyclopentyl2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-cyclopropylpropanoate

Cyclopentanol (2 mL, large excess) was added to(R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-cyclopropylpropanoicacid (250 mg, 0.711 mmol). The mixture was stirred and treated withN-Ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) (211 mg, 0.854mmol). The reaction mixture was stirred at room temperature for 24hours. The reaction mixture was partitioned between ethyl acetate (10mL) and saturated NaHCO₃ solution (10 mL). The organic phase wasseparated, washed with water and brine. Dried and evaporated. Theresidue was chromatographed [10-30% ethyl acetate/hexane] to give thetitle compound (258 mg, 0.615 mmol, 86% yield) as a colourless oil. LCMS(System A): t_(RET)=1.45 min; MH⁺ 420.

Intermediate 61: (S)-cyclopentyl 2-aminopropanoate hydrochloride

(S)-cyclopentyl 2-((tert-butoxycarbonyl)amino)propanoate (for apreparation see Intermediate 62, 4.145 g, 16.11 mmol) was dissolved in1,4-dioxane (25 mL) and then 4M hydrogen chloride in dioxane (25 mL, 100mmol) was added to the mixture and the flask was stirred at 25° C. for18h. The solvent was removed in vacuo. The product was suspended inethyl acetate and the solvent evaporated (×4) until having a white solididentified as the title compound (2.801 g, 14.46 mmol, 90% yield). ¹HNMR (400 MHz, methanol-d₄) δ ppm 5.28-5.34 (m, 1H), 4.00-4.08 (q, J=8Hz, 1H), 1.90-2.00 (m, 1H), 1.64-1.84 (m, 6H), 1.52 (d, J=8 Hz, 3H).

Intermediate 62: (S)-cyclopentyl2-((tert-butoxycarbonyl)amino)propanoate

(S)-2-((tert-butoxycarbonyl)amino)propanoic acid (5 g, 26.4 mmol) andcyclopentanol (24.01 mL, 264 mmol) was dissolved in toluene (250 mL).Then 2-(tributylphosphoranylidene)acetonitrile (14.46 mL, 52.9 mmol) wasadded to the reaction mixture and refluxed for 60 hours. The reactionmixture was cooled to room temperature. Then 100 mL of ethyl acetate wasadded to the reaction mixture. Then it was washed, first with 250 mL ofa solution saturated sodium bicarbonate, and in a second time with 250mL of brine. The organic layer was dried over MgSO₄ and afterward thesolvent removed in vacuo. The solid was then dissolved in MeOH and ranthrough a 100 g SCX column. The column were washed first with methanol,and then with NH₃ in MeOH. According to the TLC, we found out that thedesired product was in the first methanolic phase. The solvent wasremoved to get a pale yellow oil, which was chromatographed [0-30% Ethylacetate in Cyclohexane]. Appropriate fractions were combined and thesolvent removed to give the title compound (4.145 g, 16.11 mmol, 61.0%yield) as a pale yellow oil. ¹H NMR (400 MHz, methanol-d₄) δ ppm5.13-5.19 (m, 1H), 4.00-4.13 (m, 1H), 1.80-1.90 (m, 5H), 1.43 (s, 9H),1.31 (d, J=8 Hz, 3H).

Intermediate 63: (S)-(S)-1-methoxypropan-2-yl 2-amino-3-methylbutanoatehydrochloride

(S)-(S)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate (for a preparation seeIntermediate 64, 1.2 g, 4.15 mmol) was dissolved in ethyl acetate (2mL), the solution was treated with 4M hydrogen chloride in dioxan (2mL). The reaction mixture was stirred at room temperature overnight. Thesolvent was evaporated to give the title compound (725 mg, 3.21 mmol,77% yield) as a colourless gum. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.80 (brs, 3H), 5.19-5.27 (m, 1H), 3.39-3.53 (m, 2H), 3.33 (s, 3H), 2.43-2.53(m, 1H), 1.28 (d, J=8 Hz, 3H), 1.13-1.18 (m, 6H).

Intermediate 64: (S)-(S)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate

A mixture of (S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanoic acid(1.0 g, 4.60 mmol), diisopropylethylamine (1.19 g, 1.608 mL, 9.21 mmol),1-hydroxybenzotriazole hydrate (846 mg, 5.52 mmol), EDC (1.06 g, 5.53mmol), and (S)-1-methoxypropan-2-ol (1.037 g, 1.127 mL, 11.5 mmol) inDMF (5 mL) was stirred at room temperature overnight. The reactionmixture was partitioned between ethyl acetate (15 mL) and saturatedNaHCO₃ solution (15 mL). The organic phase was washed with 1Mhydrochloric acid (15 mL), water (15 mL) and brine (15 mL). The organicphase was dried and evaporated to give the title compound (1.23 g, 4.25mmol, 92% yield), as a colourless oil. ¹H NMR (400 MHz, CDCl₃) δ ppm5.13-5.19 (m, 1H), 5.06-5.10 (m, 1H (NH?)), 4.19-4.23 (m, 1H), 3.36-3.46(m, 2H), 3.33 (s, 3H), 2.10-2.18 (m, 1H), 1.45 (s, 9H), 1.23-1.27 (m,3H), 0.88-1.02 (m, 6H).

Intermediate 65: (S)-isopropyl 2-amino-3-methylbutanoate,4-methylbenzenesulphonate

(S)-2-amino-3-methylbutanoic acid (2.5 g, 21.34 mmol), tosic acid (5.28g, 27.7 mmol) and propan-2-ol (15 mL, 196 mmol) were dissolved incyclohexane (100 mL) and heated to 130° C. for 22 hrs. The solution wasallowed to cool to room temperature at which point a white solidprecipitated. This mixture was filtered under vacuum and the solidwashed several times with hexane. Solid was then placed in a vacuum ovenat 40° C. for 3 hours. This solid was then added to cyclohexane (100 mL)along with tosic acid (1.76 g, 9.2 mmol) and propan-2-ol (3.94 g, 65.3mmol). Mixture was then heated to 130° C. for 20 hours. Mixture wasallowed to cool to room temperature and filtered under gravity. Whitesolid was then placed in a vacuum oven at 40° C. for 24 hours to givethe title compound (5.977 g, 18.09 mmol, 85% yield). ¹H NMR (400 MHz,DMSO-d₆) δ ppm 8.24 (br s, 3H), 7.48 (d, J=12 Hz, 2H), 7.11 (d, J=12 Hz,2H), 4.99-5.09 (m, 1H), 3.85-3.91 (m, 1H), 2.28 (s, 3H), 2.09-2.19 (m,1H), 1.23-1.28 (m, 6H), 0.94-1.00 (m, 6H).

Intermediate 66: (S)-isobutyl 2-amino-3-methoxypropanoate, hydrochloride

10% Palladium on carbon, 50% water paste (350 mg, 20% wt) was added to astirred solution of (S)-isobutyl2-(((benzyloxy)carbonyl)amino)-3-methoxypropanoate (for a preparationsee Intermediate 67, 1.75 g, 5.66 mmol) and ammonium formate (1.78 g,28.3 mmol) in isopropanol (50 mL). The reaction mixture was stirred at90° C. for 2 hours. The reaction mixture was cooled to room temperatureand filtered through ‘celite’. The solvent was evaporated from thefiltrate. The residue was dissolved in ethyl acetate (10 mL) and treatedwith 1M hydrogen chloride in diethyl ether (6.0 mL, 6 mmol). The solventwas evaporated and the residue was triturated with diethyl ether to givethe title compound (840 mg, 3.97 mmol, 70.1% yield), as a pale yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.68 (br s, 3H), 4.32 (br s, 1H),4.01-4.06 (m, 1H), 3.88-3.94 (m, 1H), 1.92 (heptet, J=8 Hz, 1H), 0.91(d, J=8 Hz, 6H).

Intermediate 67: (S)-isobutyl2-(((benzyloxy)carbonyl)amino)-3-methoxypropanoate

A mixture of (S)-2-(((benzyloxy)carbonyl)amino)-3-methoxypropanoic acid(1.6 g, 6.32 mmol), N-ethylmorpholine (1.45 g, 1.60 mL, 12.64 mmol),N-hydoxybenzotriazole hydrate (1.16 g, 7.58 mmol), EDC (1.45 g, 7.58mmol) and isobutanol (2.34 g, 2.92 mL, 31.6 mmol) in dichloromethane (20mL) was stirred at room temperature overnight. The solvent wasevaporated. The residue was partitioned between ethyl acetate (25 mL)and saturated NaHCO₃ (25 mL). The organic phase was separated, washedwith 2M hydrochloric acid, water and brine. The organic phase was driedand evaporated. The residue was azeotroped with toluene to give thetitle compound (1.81 g, 5.85 mmol, 93% yield), as a colourless oil. ¹HNMR (400 MHz, CDCl₃) δ ppm 7.15-7.40 (m, 5H), 5.6-5.65 (m, 1H, (NH?)),5.14 (s, 2H), 4.47-4.52 (m, 1H), 3.89-4.04 (m, 2H), 3.81-3.87 (m, 1H),3.61-3.68 (m, 1H), 1.96 (heptet, J=8 Hz, 1H), 0.94 (d, J=8 Hz, 6H).

Intermediate 68: (S)-2-(((benzyloxy)carbonyl)amino)-3-methoxypropanoicacid Stage i). Ref: Tetrahedron Asymmetry 9 (1988) 3841.

A mixture of Z-Ser-OH (1.0 g, 4.18 mmol), silver(I) oxide (4.84 g, 20.9mmol) and iodomethane (5.93 g, 2.61 mL, 41.8 mmol) in acetonitrile (20mL) was stirred at room temperature overnight. The reaction mixture wasfiltered through ‘celite’ and the solvent was evaporated from thefiltrate to give stage i) product as a light yellow oil.

Stage ii).

Stage i) product was dissolved in tetrahydrofuran (5 mL) and methanol (5mL). The solution was treated with 1M lithium hydroxide solution (10mL). The reaction mixture was stirred at room temperature for 24 hours.The methanol and THF were evaporated. The residue was diluted with water(10 mL) and acidified with 2M hydrochloric acid. The mixture wasextracted with ethyl acetate (3×20 mL). The combined extracts were driedand evaporated to give the title compound (869 mg, 3.43 mmol, 82%yield), as a colourless gum. LCMS (System A): t_(RET)=0.75 min; MH⁺ 254.

Intermediate 69: (S)-isopropyl 2-amino-4-ch lorobutanoate

Thionyl chloride (2.0 g, 1.23 mL, 16.8 mmol) was added slowly to astirred solution of L-homoserine (1.0 g, 8.4 mmol) in isopropanol (15mL). The reaction mixture was stirred at 60° C. overnight. The reactionmixture was cooled and the solvent was evaporated. The residue wastriturated with diethyl ether to give a mixture. The solid waspartitioned between ethyl acetate (25 mL) and saturated NaHCO₃ (25 mL).The aqueous layer was extracted with ethyl acetate (2×25 mL). Thecombined organics were dried and evaporated to give the title compound(160 mg, 0.891 mmol, 10.61% yield), as a colourless oil. ¹H NMR (400MHz, CDCl₃) δ ppm 5.03 (heptet, J=8 Hz, 1H), 3.63-3.77 (m, 2H),3.55-3.59 (m, 1H), 2.14-2.23 (m, 1H) 1.84-1.95 (m, 1H), 1.25 (d, J=8 Hz,3H), 1.23 (d, J=8 Hz, 3H).

Intermediate 70: (4-(Ethylamino)-3-nitrophenyl)methanol

(4-fluoro-3-nitrophenyl)methanol (500 mg, 2.92 mmol), 70% ethanamine inwater (286 μl, 2.92 mmol) and DIPEA (1531 μl, 8.77 mmol) were dissolvedin tetrahydrofuran (THF) (3 mL) and the reaction mixture was heated in aBiotage Initiator microwave using initial high absorption setting to120° C. for 3 hours. The reaction mixture was partitioned between DCM(25 mL) and saturated sodium hydrogen carbonate solution (25 mL). Thelayers were separated and the aqueous layer was extracted with DCM (2×25mL). The organic layers were combined, dried using a hydrophobic fritand evaporated under reduced pressure. The crude sample was loaded indichloromethane and purified by silica gel column chromatography (50 gsilica) using a gradient of 0-5% dichloromethane-2M ammonia in methanolover 10 column volumes. The appropriate fractions were combined andevaporated under reduced pressure to give the title compound(4-(ethylamino)-3-nitrophenyl)methanol (524.3 mg, 2.67 mmol, 91% yield)as an orange solid. LCMS (System A): t_(RET)=0.80 min; MH⁺ 197. ¹H NMR δ(400 MHz, D6-DMSO): 1.22, (t, J=7 Hz, 3H), 3.34-3.42, (m, 2H), 4.39, (d,J=5 Hz, 2H), 5.18, (t, J=6 Hz, H), 7.03, (d, J=9 Hz, H), 7.48, (dd, J=6Hz, J=3 Hz, H), 7.98-8.01, (m, H), 8.02-8.07, (m, H).

The following Intermediates were prepared in a similar manner toIntermediate 70 using the appropriate commercially available amines:

(In the tables, details of the LCMS system used, retention time(t_(R)ET), MH⁺, reaction yield and % yield are provided for eachIntermediate).

Intermediate 71: (4-(methylamino)-3-nitrophenyl)methanol

(prepared from: (4-fluoro-3-nitrophenyl)methanol (commerciallyavailable)) System A, 0.64 min, MH⁺=183; Yield: 2.25 g, 106% (>100%yield due to impurities)

Intermediate 72: (4-(isopropylamino)-3-nitrophenyl)methanol

(prepared from: (4-fluoro-3-nitrophenyl)methanol (commerciallyavailable)) System A, 0.87 min, MH⁺=211; Yield: 2.49 g, 101% (>100%yield due to trace solvent present in product)

Intermediate 73:(S)-(3-nitro-4-(((tetrahydrofuran-2-yl)methyl)amino)phenyl)methanol

(prepared from: (4-fluoro-3-nitrophenyl)methanol (commerciallyavailable)) System B, 0.80 min, MH⁺=253; Yield: 1.90 g, 92%

Intermediate 74:(R)-(3-nitro-4-(((tetrahydrofuran-2-yl)methyl)amino)phenyl)methanol

(prepared from: (4-fluoro-3-nitrophenyl)methanol (commerciallyavailable)) System B, 0.80 min, MH⁺=253; Yield: 1.99 g, 96%

Intermediate 75:(4-nitro-3-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)methanol

(prepared from: (3-fluoro-4-nitrophenyl)methanol (commerciallyavailable)) System B, 0.81 min, MH⁺=267 Yield; 9.42 g, 121% (Samplecontains impurities, carried through without further purification)

Intermediate 76:(rac)-(4-(((1,4-dioxan-2-yl)methyl)amino)-3-nitrophenyl)methanol

(prepared from: (4-fluoro-3-nitrophenyl)methanol (commerciallyavailable)) System B, 0.70 min, MH⁺=269 Yield: 1.96 g, 53%

Intermediate 78:(S)-(3-((1-methoxypropan-2-yl)amino)-4-nitrophenyl)methanol

(prepared from: (3-fluoro-4-nitrophenyl)methanol (commerciallyavailable)) System A, 0.80 min, MH⁺=241 Yield: 494 mg, 23%

Intermediate 79:(S)-(4-nitro-3-(((tetrahydrofuran-2-yl)methyl)amino)phenyl)methanol

(prepared from: (3-fluoro-4-nitrophenyl)methanol (commerciallyavailable)) System A, 0.87 min, MH⁺=253; Yield: 8.49 g, 126% (Samplecontains impurities, carried through without further purification)

Intermediate 80: (R)-tert-butyl3-(((5-(hydroxymethyl)-2-nitrophenyl)amino)methyl)piperidine-1-carboxylate

(prepared from: (3-fluoro-4-nitrophenyl)methanol (commerciallyavailable)) System C, 1.13 min, MH⁺=366; Yield: 4.98 g, 93%

Intermediate 81: (S)-tert-butyl3-(((5-(hydroxymethyl)-2-nitrophenyl)amino)methyl)piperidine-1-carboxylate

(prepared from: (3-fluoro-4-nitrophenyl)methanol (commerciallyavailable)) System B, 1.18 min, MH⁺=366; Yield: 2.74 g, 103% (Samplecontains impurities, carried through without further purification)

Intermediate 82: (4-((1-methoxybutan-2-yl)amino)-3-nitrophenyl)methanol

(prepared from: (4-fluoro-3-nitrophenyl)methanol (commerciallyavailable)) System A, 0.95 min, 255 Yield: 7.18 g, 97%

Intermediate 83:(4-nitro-3-(((tetrahydrofuran-3-yl)methyl)amino)phenyl)methanol

(prepared from: (3-fluoro-4-nitrophenyl)methanol (commerciallyavailable)) System A, 0.79 min, MH⁺=253; Yield: 3.9 g, 88%

Intermediate 84: (S)-tert-butyl3-(((4-(hydroxymethyl)-2-nitrophenyl)amino)methyl)piperidine-1-carboxylate

DIPEA (7.65 mL, 43.8 mmol) was added to a stirred solution of4-Fluoro-3-nitrobenzyl alcohol (2.50 g, 14.61 mmol) and(3S)-3-(aminomethyl)piperidine (4.70 g, 21.91 mmol in2-Methyltetrahydrofuran (15 mL). The solution was heated to 80° C.overnight, the reaction mixture cooled to room temperature and theresulting solid partitioned between EtOAc and sat. aq. NaHCO₃. Theaqueous layer was removed and the organic layer washed (1×sat. aq.NaHCO₃, 1× brine). The organic portion was dried over MgSO₄ andevaporated in vacuo to an orange oil. The residue was dissolved in DCMand purified by silica gel chromatography eluting with cyclohexane:EtOAc(10-66%). The product containing fractions were evaporated in vacuo toan orange oil foam. The oil was dissolved in TBME and evaporated invacuo to an orange oil to give the title compound as an orange oil (5.52g). The total yield of the reaction was 88%. LCMS (System C):t_(RET)=1.27 min, MH⁺=366.

The following Intermediates were prepared in a similar manner toIntermediate 84 using the appropriate commercially available amines:

Intermediate 85:(4-nitro-3-((tetrahydro-2H-pyran-4-yl)amino)phenyl)methanol

(prepared from: (3-fluoro-4-nitrophenyl)methanol (commerciallyavailable)) System A, 0.80 min, MH⁺=253, Yield: 1.19 g, 77%

Intermediate 86:(4-nitro-3-(((tetrahydro-2H-pyran-2-yl)methyl)amino)phenyl)methanol

(prepared from: (3-fluoro-4-nitrophenyl)methanol (commerciallyavailable)) System A, 0.97 min, MH⁺=267 Yield: 1.41 g, 93%

Intermediate 87:(3-nitro-4-((1-(tetrahydro-2H-pyran-4-yl)ethyl)amino)phenyl)methanol

(prepared from: (3-fluoro-4-nitrophenyl)methanol (commerciallyavailable)) System A, 0.89 min, MH⁺=281 Yield: 1.06 g, 75%

Intermediate 88: (R)-tert-butyl3-(((4-(hydroxymethyl)-2-nitrophenyl)amino)methyl)piperidine-1-carboxylate

(prepared from: (3-fluoro-4-nitrophenyl)methanol (commerciallyavailable)) System A, 1.14 min, MH⁺=366; Yield: 4.41 g, 78%

Intermediate 89:5-(1-ethyl-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

4-(ethylamino)-(3-nitrophenyl)methanol (For a preparation seeIntermediate 70, 520 mg, 2.65 mmol),5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (For a preparation seeIntermediate 1, 363 mg, 2.65 mmol) and sodium dithionite (1384 mg, 7.95mmol) were added to Ethanol (8 mL) and Water (4 mL). The reactionmixture was heated in a microwave to 100° C. for 5 hours. The reactionmixture was partitioned between 25% propan-2-ol in DCM solution (25 mL)and saturated sodium hydrogen carbonate solution (25 mL) and the layerswere separated. The aqueous layer was extracted with 25% propan-2-ol inDCM solution (3×25 mL). The organic layers were combined, dried using ahydrophobic frit and evaporated under reduced pressure. The sample wasloaded in DCM and purified by silica gel column chromatography (50 gsilica) using a gradient of 2-12% DCM-2M ammonia in methanol over 10column volumes followed by holding at 12% DCM-2M ammonia in methanol for5 column volumes. The appropriate fractions were combined and evaporatedunder reduced pressure to give the title compound (295.1 mg) as a whitesolid. LCMS (System B): t_(RET)=060 min, MH⁺=284.

The following Intermediates were prepared in a similar way toIntermediate 89, using either5-Methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (Intermediate 1) or1,5-Dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (Intermediate 2)as appropriate:

Intermediate 90:5-(5-(hydroxymethyl)-1-methyl-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(prepared from: Intermediate 71) System B, 0.59 min, 284 Yield: 0.7 g,36%

Intermediate 91:5-(5-(hydroxymethyl)-1-isopropyl-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(prepared from: Intermediate 72) System A, 0.52 min, MH⁺=312; Yield: 722mg, 33%

Intermediate 92:(S)-5-(5-(hydroxymethyl)-1-((tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

(prepared from: Intermediate 73) System B, 0.63 min, MH⁺=340; Yield: 941mg, 37%

Intermediate 93:(R)-5-(5-(hydroxymethyl)-1-((tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

(prepared from: Intermediate 74) System B, 0.63 min, MH⁺=340; Yield: 168mg, 6.3%

Intermediate 94:5-(6-(hydroxymethyl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(prepared from: Intermediate 75) System B, 0.61 min, MH⁺=368; Yield: 4.5g, 95%

Intermediate 95:(rac)-5-(1-((1,4-dioxan-2-yl)methyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

(prepared from: Intermediate 76) System C, 0.44 min, MH⁺=356; Yield: 1.2g, 30%

Intermediate 97:(S)-5-(6-(hydroxymethyl)-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(prepared from: Intermediate 78) System B, 0.67 min, MH⁺=342; Yield:1.85 g, 55%

Intermediate 98:(S)-5-(6-(hydroxymethyl)-1-((tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(prepared from: Intermediate 79) System B, 0.67 min, MH⁺=354; Yield: 2.3g, 58%

Intermediate 99: (R)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate

(prepared from: Intermediate 80) System C, 0.66 min, MH⁺=467; Yield:1.78 g, 56%

Intermediate 100: (S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate

(prepared from: Intermediate 81) System A, 0.69 min, MH⁺=467; Yield: 811mg, 19%

Intermediate 101:5-(5-(hydroxymethyl)-1-(1-methoxybutan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(prepared from: Intermediate 82) System A, 0.55 min, MH⁺=356; Yield:2.76 g, 28%

Intermediate 102:5-(6-(hydroxymethyl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(prepared from: Intermediate 83) System B, 0.64 min, MH⁺=354; Yield:1.88 g, 34%

Intermediate 103: (S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate

Sodium hydrosulfite (7.89 g, 45.3 mmol) was added to a suspension of(S)-tert-butyl3-(((4-(hydroxymethyl)-2-nitrophenyl)amino)methyl)piperidine-1-carboxylate(For an example preparation see Intermediate 84) (5.52 g, 15.11 mmol)and 1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for apreparation see Intermediate 2, 2.97 g, 19.64 mmol) in a mixture ofwater (20 mL) and ethanol (40 mL). The resulting suspension was heatedto 80° C. for 6 h, cooled to room temperature and left for 1.5 days. Themixture was partitioned between sat. aq. NaHCO₃ (200 mL) and DCM:IPA(3:1, 200 mL). The organic layer was removed and the aqueous layerextracted [2×DCM:IPA (3:1, 200 mL)]. The combined organic layers weredried over MgSO₄ and evaporated in vacuo to a pale brown foam. Theresidue was dissolved in DCM, purified by silica gel chromatographyeluting with EtOAc:EtOH (12.5-25%). The product containing fractionswere evaporated in vacuo to a pale brown foam and slurried with TBME andcyclohexane. The resulting suspension was evaporated in vacuo to givethe title compound as a white solid. The total yield of the reaction was49%. LCMS (System C): t_(RET)=0.68 min, MH⁺=467.

The following Intermediates were prepared in a similar manner toIntermediate 103:

Intermediate 104:(4-nitro-3-((tetrahydro-2H-pyran-4-yl)amino)phenyl)methanol

(prepared from: Intermediate 2 and Intermediate 85) System A, 0.44 min,MH⁺=354; Yield: 0.723 g, 40%

Intermediate 105:5-(6-(hydroxymethyl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(prepared from: Intermediate 2 and Intermediate 86) System A, 0.55 min,MH⁺=368; Yield: 0.723 g, 40%

Intermediate 106:5-(5-(hydroxymethyl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(prepared from: Intermediate 2 and Intermediate 87) System A, 0.50 min,MH⁺=382; Yield: 0.416 g, 27%

Intermediate 107:5-(1-(1,3-dimethoxypropan-2-yl)-6-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(prepared from: Intermediate 2 and Intermediate 180) System B, 0.67 min,MH⁺=372; Yield: 2.038 g, 29%

Intermediate 108: (R)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate

(prepared from: Intermediate 2 and Intermediate 88) System A, 0.71 min,MH⁺=467; Yield: 2.96 g, 50%

Intermediate 109:(R)-5-(5-(hydroxymethyl)-1-(piperidin-3-ylmethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

A solution of 5M HCl in IPA (40 mL, 200 mmol) was added to(S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate(for a preparation see Intermediate 103, 1.8 g, 3.86 mmol) and thesuspension stirred for 4 h. The reaction mixture was evaporated in vacuoto a brown oil. The residue was dissolved in MeOH and loaded on to a 20g SCX cartridge. The cartridge was eluted with MeOH (200 mL), followedby 2M methanolic ammonia (100 mL). The basic fractions were evaporatedin vacuo to give the title compound as a pale yellow foam (1.389 g). Thetotal yield of the reaction was 98%. LCMS (System B): t_(RET)=0.61 min,MH⁺=367.

The following Intermediate was prepared in a similar manner toIntermediate 109:

Intermediate 110:(S)-5-(5-(hydroxymethyl)-1-(piperidin-3-ylmethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one,Hydrochloride

(prepared from: Intermediate 108) System A, 0.60 min, MH⁺=367; Yield:1.4 g, 97%

Intermediate 111:(S)-5-(1-((1-acetylpiperidin-3-yl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

Acetic anhydride (0.382 mL, 4.05 mmol) was added to a suspension of(R)-5-(5-(hydroxymethyl)-1-(piperidin-3-ylmethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 109, 1.35 g, 3.68 mmol) in2-Methyltetrahydrofuran (30 mL). The resulting suspension was stirredfor 2h. The reaction mixture was partitioned between 2-MeTHF (200 mL)and sat. aq. NaHCO₃ (25 mL). The organic layer was washed (1×sat. aq.NaHCO₃[25 mL]), dried over MgSO₄ and evaporated in vacuo to a whitesolid (˜0.3 g). The combined aqueous layers were extracted (3×DCM [50mL]) and the organic layers added to the residue from the 2-MeTHFevaporation. The resulting solution was evaporated in vacuo to a whitesolid. The solid was dissolved in DCM, and purified by silica gelchromatography eluting with DCM:2M methanolic ammonia (0-5%). Theproduct containing fractions were evaporated in vacuo to a give thetitle compound as a white solid (1.353 g). The total yield was 90%. LCMS(System C): t_(RET)=0.44 min, MH⁺=409.

Intermediate 112:(R)-5-(1-((1-acetylpiperidin-3-yl)methyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

To a stirred suspension of(S)-5-(5-(hydroxymethyl)-1-(piperidin-3-ylmethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-onehydrochloride (for a preparation see Intermediate 109, 1.49 g, 3.37mmol) in dichloromethane (10 mL) was added DIPEA (3.53 mL, 20.19 mmol).To the resulting yellow solution was added acetyl chloride (0.718 mL,10.10 mmol) and stirred for 1.5 h. 2M aq. NaOH (10 mL, 20.00 mmol) wasadded, the reaction mixture stirred vigorously for 30 min, and theorganic layer removed. The aqueous was extracted 3 times with DCM andthe combined organic phases passed through a hydrophobic frit andconcentrated in vacuo to give a brown paste. The residue was dissolvedin THF (10 mL) and 2M aq. NaOH (10 mL, 20.00 mmol) was added. Thereaction mixture was stirred for 30 min. The reaction mixture waspartitioned between Water and DCM and the organic layer removed. Theaqueous phases was extracted 3 times with DCM. Sat. aq. NaHCO₃ andDCM:IPA (3:1) were added to the aqueous layer and the organic removed.The aqueous layer was extracted 3 times and the combined organic phaseswere passed through a hydrophobic frit, concentrated and dried in vacuoto give a yellow solid. The crude solid was dissolved in DCM, purifiedby silica gel chromatography, eluting with 2M Methanolic ammonia:DCM(2.5-12.5%, 15 CV). The appropriate fractions were combined andevaporated in vacuo to give the title compound as a yellow solid (1.01g). The total yield of the reaction was 73%. LCMS (System A):t_(RET)=0.47 min, MH⁺=409.

Intermediate 113:1-ethyl-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde

5-(1-ethyl-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(For a preparation see Intermediate 89, 290 mg, 1.024 mmol) and 45% byweight 2-iodoxybenzoic acid (701 mg, 1.126 mmol) were added to DCM (5mL) and the suspension was stirred under nitrogen for 4 days. Thereaction mixture was partitioned between saturated sodium hydrogencarbonate solution (30 mL) and DCM (30 mL) and the layers wereseparated. The aqueous layer was extracted with DCM (3×30 mL). Theorganic layers were combined and concentrated under reduced pressure. Itwas again partitioned between saturated sodium hydrogen carbonatesolution (50 mL) and DCM (50 mL). The layers were separated and theaqueous layer was extracted with DCM (3×50 mL). The combined organiclayers were dried using a hydrophobic frit and was concentrated underreduced pressure to give the title compound (263 mg) as an off-whitesolid. LCMS (System B): t_(RET)=0.67 min, MH⁺=282

The following Intermediates were prepared in a similar way toIntermediate 113:

Intermediate 114:(S)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

(prepared from: Intermediate 92(S)-5-(5-(hydroxymethyl)-1-((tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one)System B, 0.70 min, MH⁺=338; Yield: 1.04 g, 85%

Intermediate 115:(R)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

(prepared from: Intermediate 93) System B, 0.70 min, MH⁺=338; Yield: 147mg, 63%

Intermediate 117:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-benzo[d]imidazole-5-carbaldehyde

(prepared from: Intermediate 101) System A, 0.82 min, MH⁺=354; Yield:2.60 g, 95%

Intermediate 118:(S)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

A suspension of(S)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(For a preparation see Intermediate 114, 500 mg, 1.260 mmol) andPotassium carbonate (348 mg, 2.52 mmol) in DMF (5 mL) was stirred for1.15 h prior to adding iodomethane (0.095 mL, 1.512 mmol). The reactionmixture was stirred over the weekend. The reaction mixture wasconcentrated under reduced pressure and the residue was partitionedbetween 3:1 chloroform:isopropanol (125 mL) and saturated solution ofsodium bicarbonate (125 mL). The organic layer was isolated and theaqueous fraction was re-extracted three times with 3:1chloroform:isopropanol (3×125 mL). The organic fractions were combined,passed through a hydrophobic frit and concentrated under reducedpressure. The residue was dissolved in 10% ethanol in ethyl acetate andloaded onto a silica column (50 g). The products were eluted with agradient of 0-30% ethanol in ethyl acetate. The appropriate fractionswere combined and concentrated under reduced pressure to give the titlecompound (121 mg) as a light brown solid. LCMS (System B): t_(RET)=0.75min, MH⁺=352

Intermediate 119:(R)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

(R)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(For a preparation see Intermediate 115, 149 mg, 0.353 mmol) wasdissolved in DMF (2.5 mL) and Potassium carbonate (98 mg, 0.707 mmol)was added. The reaction mixture was stirred for one hour prior to addingIodomethane (0.027 mL, 0.424 mmol). The flask containing the reactionmixture was sealed. The reaction mixture was stirred overnight. Thereaction mixture was diluted with isopropanol and filtered through acelite cartridge (2.5 g) which had been preconditioned with the solvent.4 CV of isopropanol were passed through the column. The washings werecombined and concentrated under reduced pressure. The residue wasdissolved in a minimum amount of DCM and loaded onto a silica column (25g) and eluted with a gradient of 0-20% ethanol in ethyl acetate. Theappropriate fractions were combined and concentrated under reducedpressure to give the title compound (56 mg) as a colourless gum. LCMS(System B): t_(RET)=0.75 min, MH⁺=352

Intermediate 120:1-((1,4-dioxan-2-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde(Single Enantiomer of Unknown Configuration)

Potassium carbonate (368 mg, 2.66 mmol) was added in a single portion toa suspension of1-((1,4-dioxan-2-yl)methyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde(For a preparation see Intermediate 122, 470 mg, 1.330 mmol) in DMF (15mL). The mixture was then cooled in a water-ice bath and iodomethane(0.108 mL, 1.729 mmol) was added dropwise. When the addition wascompleted, the bath was removed and the reaction mixture stirred at rtovernight (18 hr). The solvent was removed under reduced pressure. Thesolid was then partitioned between NaHCO₃ (100 mL) and DCM:iPrOH 3:1(100 mL). The separated aqueous phase was extracted with DCM:iPrOH 3:1(3×100 mL). The combined organic phases where passed through ahydrophobic frit and evaporated to obtain the title compound (476 mg) asa pale yellow solid. LCMS (System B): t_(RET)=0.70 min, MH⁺=368.

Intermediate 121:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazole-6-carbaldehyde

Manganese dioxide (11 g, 108 mmol) was added in a single portion to astirred solution of5-(6-(hydroxymethyl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(For a preparation see Intermediate 94, 3.3 g, 8.98 mmol) in chloroform(150 mL) at rt. The resultant suspension was stirred rapidly overnight(18 hr). Further manganese dioxide (2 g, 19.55 mmol) was added and thereaction mixture was stirred for 0.5 hr longer. 20 mL of the reactionmixture was filtrated through celite and the solvent evaporated toobtain the title compound (424 mg) as colourless oil. LCMS (System B):t_(RET)=0.72 min, MH⁺=367

Intermediate 122:1-((1,4-dioxan-2-yl)methyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde(Single Unknown Enantiomer)

Manganese dioxide (2232 mg, 21.82 mmol) was added in a single portion toa stirred suspension of5-(1-((1,4-dioxan-2-yl)methyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(For a preparation see Intermediate 138a, 554 mg, 1.559 mmol) inchloroform (25 mL) at rt. The resultant suspension was stirred rapidlyfor 4 hr and was left to stand overnight. The suspension was filteredthrough Celite and flushed with DCM (2×30 mL), MeOH (5×30 mL) andDCM:iPr 3:1 (2×30 mL). The filtrate was evaporated under vacuo and 1 hrin the vacuum oven to obtain the title compound (470 mg) as a paleyellow solid. LCMS (System B): t_(RET)=0.65 min, MH⁺=354

The following Intermediates were prepared in a similar way toIntermediate 122:

Intermediate 123:(S)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazole-6-carbaldehyde

(prepared from: Intermediate 97) System A, 0.71 min, MH⁺=340; Yield:1.56 g, 87%

Intermediate 124:(S)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazole-6-carbaldehyde

(prepared from: Intermediate 98) System B, 0.80 min, MH⁺=352; Yield:2.29 g, 100%

Intermediate 125:(R)-1-((1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-6-carbaldehyde

(prepared from: Intermediate 141) System B, 0.73 min, MH⁺=407; Yield:535 mg, 79%

Intermediate 126: (R)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-formyl-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate

(prepared from: Intermediate 99) System C, 0.80 min, MH⁺=465; Yield: 897mg, 90%

Intermediate 127:(S)-1-((1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-6-carbaldehyde

(prepared from: Intermediate 142) System B, 0.73 min, MH⁺=407; Yield:217 mg, 95%

Intermediate 128: (S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-formyl-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate

(prepared from: Intermediate 100) System B, 1.00 min, MH⁺=465; Yield: 79mg, 64%

Intermediate 129:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazole-6-carbaldehyde

(prepared from: Intermediate 102) System B, 0.74 min, MH⁺=352; Yield:1.79 g, 96%

Intermediate 130: (S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-5-formyl-1H-benzo[d]imidazole-1-yl)methyl)piperidine-1-carboxylate

Manganese dioxide (2.98 g, 34.3 mmol) was added to a solution of(S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-5-(hydroxymethyl)-1Hbenzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate (1.6 g, 3.43 mmol)(for an example preparation see Intermediate 103) in chloroform (60 mL).The suspension was stirred for 3 h and stirring stopped over theweekend. The suspension was filtered and evaporated in vacuo to give thetitle compound as a brown solid (1.299 g). The total yield for thereaction was 82%. LCMS (System C): t_(RET)=0.83 min, MH⁺=465.

The following Intermediates were prepared in a similar manner toIntermediate 130:

Intermediate 131:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazole-6-carbaldehyde

(prepared from: Intermediate 104) System A, 0.67 min, MH⁺=351 Yield:0.657 g, 87%

Intermediate 132:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazole-6-carbaldehyde

(prepared from: Intermediate 105) System A, 0.84 min, MH⁺=366; Yield:0.983 g, 94%

Intermediate 133:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-benzo[d]imidazole-5-carbaldehyde

(prepared from: Intermediate 106) System A, 0.74 min, MH⁺=380; Yield:0.376 g, 88%

Intermediate 134:1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-6-carbaldehyde

(prepared from: Intermediate 107) System B, 0.80 min, MH⁺=370; yield notrecorded)

Intermediate 135:(S)-1-((1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde

(prepared from: Intermediate 111) System C, 0.55 min, MH⁺=407; Yield:1.41 g, 94%

Intermediate 136:(R)-1-((1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde

(prepared from: Intermediate 112) System A, 0.65 min, MH⁺=407; Yield:958 mg, 91%

Intermediate 137: (2S,3R)-Cyclopentyl 2-amino-3-hydroxybutanoate,4-methylbenzenesulphonic acid salt

To a suspension of (2S,3R)-2-amino-3-hydroxybutanoic acid (20 g, 168mmol) in cyclohexane (200 mL), cyclopentanol (116 g, 1343 mmol) and4-methylbenzenesulfonic acid (37.6 g, 218 mmol) at room temperature wereadded. The reaction mixture was stirred at 100° C. for 24 hr. Thereaction mixture was evaporated in vacuo to give the crude product as abrown oil. The brown oil was allowed to cool and the resulting crystalsfiltered, washed with EtOAc (50 mL) to give (2S,3R)-cyclopentyl2-amino-3-hydroxybutanoate, 4-methylbenzenesulphonic acid salt (50.06 g,136 mmol, 81% yield) as a white solid. ¹H NMR δ (400 MHz, DMSO-d₆) ppm:8.20 (3H, br. s.), 7.48 (2H, d, J=8.1 Hz), 7.12 (2H, d, J=7.8 Hz), 5.63(1H, d, J=4.4 Hz), 5.20 (1H, t, J=5.6 Hz), 4.18-4.03 (1H, m), 3.89 (1H,d, J=3.4 Hz), 2.30 (3H, s), 1.94-1.78 (2H, m), 1.77-1.50 (6H, m), 1.20(3H, d, J=6.6 Hz).

Intermediate 138a and 138b:5-(1-((1,4-dioxan-2-yl)methyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

(rac)-5-(1-((1,4-dioxan-2-yl)methyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(for a preparation see Intermediate 95, 1.2 g) was separated into it'stwo corresponding enantiomers by chiral chromatography, using a 30 mm×25cm Chiralcel OD-H column, eluting with 40% EtOH/Heptane at a flowrate of30 mL/min, wavelength of detection=215 nm.

Isomer 1: (Intermediate 138a) 563 mg Obtained as a Solid

LCMS (System A): t_(RET)=0.44 min, MH⁺=356

Analysed for chiral purity on 4.6 mmid×25 cm Chiralcel OD-H column,eluting with 40% EtOH/Heptane at a flowrate of 1.0 mL/min, wavelength ofdetection=215 nm.

Chiral purity found to be >99.5%.

Isomer 2: (Intermediate 138b) 598 mg Obtained as a Solid

LCMS (System A): t_(RET)=0.41 min, MH⁺=356

Analysed for chiral purity on 4.6 mmid×25 cm Chiralcel OD-H column,eluting with 40% EtOH/Heptane at a flowrate of 1.0 mL/min, wavelength ofdetection=215 nm.

Chiral purity found to be 99.1%.

Intermediate 139:(S)-5-(6-(hydroxymethyl)-1-(piperidin-3-ylmethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

5M HCl in IPA (15 mL) was added to (R)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate(For a preparation see Intermediate 99, 850 mg, 1.82 mmol). Theresulting solution was stirred for 3 h and evaporated in vacco to abrown solid. The residue was dissolved in MeOH, loaded on to a 10 g SCXcartridge and eluted with MeOH, followed by 2M methanolic ammonia. Thebasic fractions were evaporated in vacuo to give the title compound (617mg) as a white foam. LCMS (System C): t_(RET)=0.33 min, MH⁺=367.

Intermediate 140:(R)-(1-((1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-6-yl)methylacetate

(S)-5-(6-(hydroxymethyl)-1-(piperidin-3-ylmethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(For a preparation see Intermediate 139, 617 mg, 1.68 mmol) wasdissolved in DCM (5 mL). DIPEA (0.618 mL, 3.54 mmol) and acetyl chloride(0.253 mL, 3.54 mmol) were added and the reaction stirred under nitrogenat rt for 3.5 hr. 0.2 eq of DIPEA (0.059 mL, 0.336 mmol) and 0.2 eq ofacetyl chloride (0.024 mL, 0.336 mmol) were added and the reaction wasstirred under nitrogen overnight (16 hr). 0.4 eq of DIPEA (0.118 mL,0.672 mmol) and 0.4 eq of acetyl chloride (0.048 mL, 0.672 mmol) wereadded and the reaction was stirred under nitrogen for 1 hr. 5 mL of aq.NaOH 2M were added and the suspension was vigorously stirred for 30 min.The separated aqueous phase was extracted with DCM (3×10 mL). Thecombined organic phases were passed through a hydrophobic frit to obtainthe title compound (796 mg) as a yellow paste. LCMS (System A):t_(RET)=0.62 min, MH⁺=451.

Intermediate 141:(R)-5-(1-((1-acetylpiperidin-3-yl)methyl)-6-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(R)-(1-((1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-6-yl)methylacetate (For a preparation see Intermediate 140, 796 mg, 1.77 mmol) wasdissolved in tetrahydrofuran (2 mL) and MeOH (2 mL) and aqueous NaOH 2M(0.883 mL, 1.77 mmol) was added. The reaction mixture was stirred for 10min at rt. The reaction mixture was neutralized to pH=7 with aq. sol.HCl 2M and diluted with Water (10 mL) and DCM:iPrOH 3:1 (10 mL). Theseparated aqueous phase was extracted with DCM:iPrOH 3:1 (3×10 mL). Thecombined organic phases were passed through a hydrophobic frit andevaporated to obtain the title compound (683 mg) as a yellow oil. LCMS(System A): t_(RET)=0.46 min, MH⁺=409.

Intermediate 142:(S)-5-(1-((1-acetylpiperidin-3-yl)methyl)-6-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(R)-5-(6-(hydroxymethyl)-1-(piperidin-3-ylmethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(For a preparation see Intermediate 170, 284 mg, 0.775 mmol) wasdissolved in DCM (5 mL) and cooled in a water/ice bath prior addition ofDIPEA (0.108 mL, 0.620 mmol) followed by dropwise addition of acetylchloride (0.044 mL, 0.620 mmol). The reaction mixture was stirred for 5min, then the bath was removed and the reaction mixture was allowed towarm up to rt and stirred for 2 hr. The reaction mixture was partitionedbetween DCM:iPrOH 3:1 (10 mL) and NaHCO₃ (10 mL). The separated aqueousphase was extracted with DCM:iPrOH 3:1 (3×10 mL). The combined organicfractions were passed though a hydrofobic frit and evaporated to obtainthe crude product, 300 mg. The samples were dissolved in MeOH 3 mL andpurified by MDAP (Method B). The solvent was dried down to give thetitle compound (230 mg) as a white solid. LCMS (System B): t_(RET)=0.64min, MH⁺=409.

Intermediate 143: (2S)-cyclopentyl4-methyl-2-((4-(((4-methylmorpholin-2-yl)methyl)amino)-3-nitrobenzyl)amino)pentanoate

(4-methylmorpholin-2-yl)methanamine (665 mg, 5.11 mmol) and DIPEA (0.892mL, 5.11 mmol) were added to a solution of (S)-cyclopentyl2-((4-fluoro-3-nitrobenzyl)amino)-4-methylpentanoate (for a preparationsee Intermediate 144, 600 mg, 1.703 mmol) in tetrahydrofuran (12 mL).The reaction mixture was heated in a Biotage Initiator microwave usinginitial high absorption setting to 120° C. for a total of 90 mins. DCM(30 mL) and saturated sodium hydrogen carbonate solution (30 mL) wereadded and the layers were separated. The aqueous layer was extractedwith DCM (2×30 mL) and the organic layers were combined, dried andevaporated under reduced pressure to give an orange liquid. The samplewas loaded in dichloromethane and purified by silica gel columnchromatography (50 g silica) using a gradient of 0-10%dichloromethane-methanol over 15 column volumes followed by holding at10% dichloromethane-methanol for 5 column volumes. The appropriatefractions were combined and evaporated under reduced pressure to givethe title compound (693.6 mg, 1.499 mmol, 88% yield) as an orange gum.LCMS (System A): t_(RET)=0.70 min, MH⁺=463

Intermediate 144: (S)-cyclopentyl2-((4-fluoro-3-nitrobenzyl)amino)-4-methylpentanoate

4-fluoro-3-nitrobenzaldehyde (2 g, 11.83 mmol) and (S)-cyclopentyl2-amino-4-methylpentanoate 4-methylbenzenesulfonate (for a preparationsee Intermediate 3, 4.83 g, 13.01 mmol) were dissolved in DCM (50 mL)and to this, acetic acid (2.031 mL, 35.5 mmol) was added. The reactionmixture was stirred under nitrogen for 1.5 hours. sodiumtriacetoxyborohydride (5.01 g, 23.65 mmol) was added in portions and thereaction mixture was stirred under nitrogen overnight. Saturated aqueoussodium hydrogen carbonate solution (100 mL) was added slowly and thereaction mixture was stirred until the fizzing had stopped. Theresulting suspension was extracted with DCM (3×100 mL). The combinedorganic layers were dried and evaporated under reduced pressure to givea yellow oil. The crude sample was loaded in dichloromethane andpurified by Biotage SP4 SNAP 100 g silica using a gradient of 0-50%cyclohexane-ethyl acetate over 10 column volumes followed by holding at50% cyclohexane-ethyl acetate for 10 column volumes. The appropriatefractions were combined and evaporated under reduced pressure to givethe title compound (2.64 g, 7.48 mmol, 63.3% yield) as a yellow oil.

LCMS (System A): t_(RET)=0.97 min, MH⁺=353.

Intermediate 145: (2S)-cyclopentyl2-((4-(((4-methylmorpholin-2-yl)methyl)amino)-3-nitrobenzyl)amino)propanoate

(S)-cyclopentyl 2-((4-fluoro-3-nitrobenzyl)amino)propanoate (for apreparation see Intermediate 146, 490 mg, 1.579 mmol),(4-methylmorpholin-2-yl)methanamine (617 mg, 4.74 mmol) and DIPEA (0.827mL, 4.74 mmol) were dissolved in THF (12 mL) and the reaction mixturewas heated in a Biotage Initiator microwave using initial highabsorption setting to 120° C. for a total of 90 minutes. The reactionmixture was partitioned between saturated sodium hydrogen carbonatesolution (40 mL) and DCM (40 mL) and the layers were separated. Theaqueous layer was washed with DCM (3×40 mL) and the combined organiclayers were dried using a hydrophobic frit and evaporated under reducedpressure to give an orange oil. The sample was loaded in dichloromethaneand purified by silica gel column chromatography (50 g silica) using agradient of 0-12% dichloromethane-2M ammonia in methanol over 10 columnvolumes followed by holding at 12% dichloromethane-2M ammonia inmethanol for 5 column volumes. The appropriate fractions were combinedand evaporated under reduced pressure to give the title compound (649.8mg, 1.545 mmol, 98% yield) as an orange oil. LCMS (System A):t_(RET)=1.12 min, MH⁺=421

Intermediate 146: (S)-cyclopentyl2-((4-fluoro-3-nitrobenzyl)amino)-4-methylpentanoate

4-fluoro-3-nitrobenzaldehyde (500 mg, 2.96 mmol) and (S)-cyclopentyl2-aminopropanoate hydrochloride (for a preparation see Intermediate 61,630 mg, 3.25 mmol) were dissolved in Dichloromethane (DCM) (15 mL) andto this solution, acetic acid (0.508 mL, 8.87 mmol) was added. Thereaction mixture was stirred under nitrogen for 1 hour. sodiumtriacetoxyborohydride (1.253 g, 5.91 mmol) was added to the reactionmixture and it was stirred under nitrogen at room temperature overnight.Saturated sodium hydrogen carbonate solution (40 mL) was slowly added tothe solution until the fizzing stopped. This solution was then extractedwith DCM (4×40 mL) and the organic layers were combined, dried using ahydrophobic frit and the solvent was removed under reduced pressure togive a yellow oil. The sample was loaded in dichloromethane and purifiedby silica gel column chromatography (50 g silica) using a gradient of35-65% cyclohexane-ethyl acetate over 10 column volumes followed byholding at 65% cyclohexane-ethyl acetate for 5 column volumes. Theappropriate fractions were combined and evaporated under reducedpressure to give the title compound (496.5 mg, 1.600 mmol, 54.1% yield)as a yellow oil. LCMS (System A): t_(RET)=0.75 min, MH⁺=311

Intermediate 147: (2S)-cyclopentyl3-methyl-2-((4-(((4-methylmorpholin-2-yl)methyl)amino)-3-nitrobenzyl)amino)butanoate

(4-methylmorpholin-2-yl)methanamine (666 mg, 5.12 mmol) and DIPEA (0.893mL, 5.12 mmol) were added to a solution of (S)-cyclopentyl2-((4-fluoro-3-nitrobenzyl)amino)-3-methylbutanoate (for a preparationsee Intermediate 148, 577 mg, 1.705 mmol) in THF (12 mL) and thereaction mixture was heated in a Biotage Initiator microwave usinginitial high absorption setting to 120° C. for a total of 90 minutes.The reaction mixture was partitioned between DCM (40 mL) and saturatedsodium hydrogen carbonate solution (40 mL) and the layers wereseparated. The aqueous layer was extracted with DCM (2×40 mL) and thecombined organic layers were dried using a hydrophobic frit andevaporated under reduced pressure to give an orange oil. The sample wasloaded in dichloromethane and purified by silica gel columnchromatography (50 g silica) using a gradient of 0-7% dichloromethane-2Mammonia in methanol over 10 column volumes followed by holding at 7%dichloromethane-2M ammonia in methanol for 5 column volumes. Theappropriate fractions were combined and evaporated under reducedpressure to give the title compound (760.6 mg, 1.696 mmol, 99% yield) asan orange oil. LCMS (System B): t_(RET)=1.33 min, MH⁺=449.

Intermediate 148: (S)-cyclopentyl2-((4-fluoro-3-nitrobenzyl)amino)-3-methylbutanoate

4-fluoro-3-nitrobenzaldehyde (500 mg, 2.96 mmol) and (S)-cyclopentyl2-amino-3-methylbutanoate 4-methylbenzenesulfonate (1.163 g, 3.25 mmol,for a preparation see Intermediate 24) were dissolved in Dichloromethane(DCM) (20 mL) and the reaction mixture was stirred under nitrogen for 1hour. Sodium triacetoxyborohydride (1.253 g, 5.91 mmol) was added andthe reaction mixture was stirred under nitrogen at room temperature for2 hours. Saturated aqueous sodium hydrogen carbonate solution (50 mL)was added to the reaction mixture in portions till the fizzing stopped.The resulting suspension was extracted with DCM (4×50 mL) and theorganic layers were combined, dried using a hydrophobic frit andevaporated under reduced pressure to give a pale yellow oil. The samplewas loaded in dichloromethane and purified by silica gel columnchromatography (50 g silica) using a gradient of 0-1%dichloromethane-methanol over 10 column volumes followed by holding at1% dichloromethane-methanol for 5 column volumes. The appropriatefractions were combined and evaporated under reduced pressure to givethe title compound (581 mg, 1.717 mmol, 35.9% yield) as a pale yellowoil. LCMS (System A): t_(RET)=0.95 min, MH⁺=339.

Intermediate 149: (S)-cyclopentyl4-methyl-2-((4-(((1-methylpiperidin-4-yl)methyl)amino)-3-nitrobenzyl)amino)pentanoate

(S)-cyclopentyl 2-((4-fluoro-3-nitrobenzyl)amino)-4-methylpentanoate(for a preparation see Intermediate 144, 500 mg, 1.419 mmol),(1-methylpiperidin-4-yl)methanamine (546 mg, 4.26 mmol) and DIPEA (0.743mL, 4.26 mmol) were added to Tetrahydrofuran (THF) (12 mL) and thereaction mixture was heated in a Biotage Initiator microwave usinginitial high absorption setting to 120° C. for 1 hour. The reactionmixture was partitioned between DCM (40 mL) and saturated sodiumhydrogen carbonate solution (40 mL) and the layers were separated. Theaqueous layer was extracted with DCM (2×40 mL) and the combined organiclayers were dried and evaporated under reduced pressure to give a yellowoil. The sample was loaded in dichloromethane and purified by silica gelcolumn chromatography (50 g silica) using a gradient of 0-10%dichloromethane-2M ammonia in methanol over 10 column volumes followedby holding at 10% dichloromethane-2M ammonia in methanol for 5 columnvolumes. The appropriate fractions were combined and evaporated underreduced pressure to give the title compound (595.5 mg, 1.293 mmol, 91%yield) as an orange gum. LCMS (System A): t_(RET)=0.69 min, MH⁺=461.

Intermediate 150:5-(5-(hydroxymethyl)-1-((4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

Three identical mixtures of(4-(((4-methylmorpholin-2-yl)methyl)amino)-3-nitrophenyl)methanol (1 g,3.55 mmol, for a preparation see Intermediate 166),5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (504 mg, 3.68 mmol,for a preparation see Intermediate 1) and sodium dithionite (2 g, 11.5mmol) in ethanol (8 mL) and water (4 mL) were heated in a BiotageInitiator microwave using initial high absorption setting to 100° C. for5 hours. The reaction mixtures were combined and partitioned betweensaturated sodium hydrogen carbonate solution (100 mL) and 25%propan-2-ol in chloroform (100 mL). The layers were separated and theaqueous layer was extracted with 25% propan-2-ol in chloroform (4×100mL). The organic layers were combined, dried using a hydrophobic fritand evaporated under reduced pressure to give a white solid. The samplewas loaded in dichloromethane and purified by Biotage SP4 SNAP 2×100 gsilica using a gradient of 5-15% dichloromethane-2M ammonia in methanolover 15 column volumes followed by holding at 15% dichloromethane-2Mammonia in methanol for 5 column volumes. The appropriate fractions werecombined and evaporated under reduced pressure to give the titlecompound5-(5-(hydroxymethyl)-1-((4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(1.745 g, 4.74 mmol, 44.4% yield) as an off-white solid. LCMS (SystemB): t_(RET)=0.58 min, MH⁺=369.

Intermediate 151:2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

5-(5-(hydroxymethyl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(for a preparation see Intermediate 152, 1.67 g, 4.73 mmol) was fullydissolved in Dimethyl Sulfoxide (DMSO) (25 mL) before adding2-iodoxybenzoic acid (3.23 g, 5.20 mmol) and stirring under nitrogen for2 hours. The reaction mixture was diluted with water (100 mL) and thewhite solid which precipitated out was removed by filtration and keptaside. The filtrate was extracted with ethyl acetate (3×100 mL) and thenwith 25% methanol in DCM solution (4×100 mL). The solid obtained fromearlier filtration was suspended in 25% methanol in DCM (100 mL) andsaturated sodium hydrogen carbonate solution (100 mL) was added. Thelayers were separated and the aqueous layer was extracted with 25%methanol in DCM solution (4×100 mL). All the organic layers werecombined, dried using a hydrophobic frit and evaporated under reducedpressure to give a white solid which was then loaded as a suspension inmethanol/dichloromethane and purified by silica gel columnchromatography (100 g silica) using a gradient of 0-10%dichloromethane-methanol over 10 column volumes followed by holding at10% dichloromethane-methanol for 10 column volumes. The pure fractionswere combined and evaporated under reduced pressure to give the titlecompound (1.1487 g, 3.27 mmol, 69.2% yield) as an off-white solid. LCMS(System B): t_(RET)=0.68 min, MH⁺=352.

Intermediate 152:5-(5-(hydroxymethyl)-1-((tetrahydro-2-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

Four identical mixtures of(3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)methanol (fora preparation see Intermediate 77, 1.15 g, 4.3 mmol),5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (592 mg, 4.3 mmol, fora preparation see intermediate 1) and sodium dithionite (2.25 g, 13.0mmol) in Ethanol (8 mL) and Water (4 mL) were heated in a BiotageInitiator microwave using initial high absorption setting to 100° C. for5 hours. All four reaction mixtures were combined and partitionedbetween saturated sodium hydrogen carbonate solution (100 mL) and 25%propan-2-ol in DCM (100 mL). The layers were separated and the aqueouslayer was extracted with 25% propan-2-ol in DCM (3×100 mL). The organiclayers were combined, dried using a hydrophobic frit and evaporatedunder reduced pressure to give a white solid. The sample was loaded indichloromethane and purified by silica gel column chromatography (2×100g silica) using a gradient of 0-10% dichloromethane-2M ammonia inmethanol over 15 column volumes followed by holding at 10%dichloromethane-2M ammonia in methanol for 10 column volumes. Theappropriate fractions were combined and evaporated under reducedpressure to give the title compound (1.8562 g, 5.25 mmol, 30.4% yield)as an off-white solid. LCMS (System A): t_(RET)=0.46 min, MH⁺=354.

Intermediate 153:1-((1-methyl-5-oxopyrrolidin-3-yl)methyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde

45% by weight 2-iodoxybenzoic acid (695 mg, 1.117 mmol) was added inportions to a suspension of5-(5-(hydroxymethyl)-1-((1-methyl-5-oxopyrrolidin-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(for a preparation see Intermediate 154, 372 mg, 1.015 mmol) in DCM (5mL) and the reaction mixture was stirred under nitrogen for 3 days. Thereaction mixture was partitioned between DCM and saturated sodiumhydrogen carbonate solution. The aqueous layer was extracted with DCM(6×40 mL). The organic layers were combined, dried using a hydrophobicfrit and evaporated under reduced pressure to give the title compound(272 mg, 0.746 mmol, 73.5% yield) as an off-white solid. LCMS (SystemB): t_(RET)=0.58 min, MH⁺=365

Intermediate 154:5-(5-(hydroxymethyl)-1-((1-methyl-5-oxopyrrolidin-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

4-(((4-(hydroxymethyl)-2-nitrophenyl)amino)methyl)-1-methylpyrrolidin-2-one(for a preparation see Intermediate 155, 710 mg, 2.54 mmol),5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparation seeintermediate 1, 349 mg, 2.54 mmol) and sodium dithionite (1328 mg, 7.63mmol) were added to ethanol (8 mL) and water (4 mL) and the reactionmixture was heated in a Biotage Initiator microwave using initial highabsorption setting to 100° C. for 5 hours. The reaction mixture waspartitioned between saturated sodium hydrogen carbonate solution (30 mL)and DCM (30 mL). The aqueous layer was extracted with DCM (3×30 mL). Theorganic layers were combined, dried using a hydrophobic frit andevaporated under reduced pressure. The sample was loaded indichloromethane and purified by silica gel column chromatography (50 gsilica) using a gradient of 5-20% dichloromethane-2M ammonia in methanolover 15 column volumes followed by holding at 20% dichloromethane-2Mammonia in methanol for 5 column volumes. The appropriate fractions werecombined and evaporated under reduced pressure to give the titlecompound (378 mg, 1.032 mmol, 40.6% yield) as a colourless gum. LCMS(System B): t_(RET)=0.51 min, MH⁺=365.

Intermediate 155:4-(((4-(hydroxymethyl)-2-nitrophenyl)amino)methyl)-1-methylpyrrolidin-2-one

(4-fluoro-3-nitrophenyl)methanol (500 mg, 2.92 mmol),4-(aminomethyl)-1-methylpyrrolidin-2-one (562 mg, 4.38 mmol) and DIPEA(1.531 mL, 8.77 mmol) were added to tetrahydrofuran (THF) (2 mL) and thereaction mixture was heated in a Biotage Initiator microwave usinginitial high absorption setting to 120° C. for 4 hours. The reactionmixture was partitioned between DCM (25 mL) and saturated sodiumhydrogen carbonate (25 mL). The layers were separated and the aqueouslayer was extracted with DCM (3×25 mL). The organic layers werecombined, dried using a hydrophobic frit and evaporated under reducedpressure. The sample was loaded in dichloromethane and purified bysilica gel column chromatography (50 g silica) using a gradient of 0-10%dichloromethane-2M ammonia in methanol over 10 column volumes followedby holding at 10% dichloromethane-2M ammonia in methanol for 5 columnvolumes. The appropriate fractions were combined and evaporated underreduced pressure to give the title compound (713.7 mg, 2.56 mmol, 87%yield) as an orange solid. LCMS (System B): t_(RET)=0.61 min, MH⁺=262.

Intermediate 156:1-((1-acetylpyrrolidin-3-yl)methyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde

5-(1-((1-acetylpyrrolidin-3-yl)methyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(For a preparation see Intermediate 157, 135 mg, 0.355 mmol) wassuspended in DCM (5 mL) and 45% by weight 2-iodoxybenzoic acid (243 mg,0.390 mmol) was added. The reaction mixture was stirred at roomtemperature for 40 hours. The reaction mixture was partitioned betweenDCM (25 mL) and saturated sodium hydrogen carbonate solution (25 mL) andthe aqueous layer was extracted with DCM (4×25 mL). The combined organiclayers were dried using hydrophobic frit and concentrated under reducedpressure. The sample was partitioned again between DCM (25 mL) andsaturated sodium hydrogen carbonate solution (25 mL) and the aqueouslayer was extracted with DCM (8×25 mL). The combined organic layers weredried using a hydrophobic frit and solvent was removed under reducedpressure to give the title compound (132.1 mg, 0.349 mmol, 98% yield) asan off-white solid. LCMS (System B): t_(RET)=0.59 min, MH⁺=264.

Intermediate 157:5-(1-((1-acetylpyrrolidin-3-yl)methyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

A solution of5-(5-(hydroxymethyl)-1-(pyrrolidin-3-ylmethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(For a preparation see Intermediate 158, 150 mg, 0.443 mmol) in DCM (5mL) and pyridine (0.108 mL, 1.330 mmol) was cooled to −5° C. and stirredfor 30 minutes. acetyl chloride (0.032 mL, 0.443 mmol), was added andthe reaction mixture was stirred at room temperature for 3 days. Thereaction mixture was partitioned between water (20 mL) and DCM (20 mL)and the aqueous layer was extracted with DCM (3×20 mL). The combinedorganic layers were dried using a hydrophobic frit and evaporated underreduced pressure to give crude material (181.8 mg). This was suspendedin THF (2 mL) and methanol (2 mL) and 1M aqueous lithium hydroxidesolution (1.29 mL, 1.29 mmol) was added and the reaction mixture wasstirred at 62° C. for 24 hours. The reaction mixture was partitionedbetween water (30 mL) and 25% propan-2-ol in DCM solution (30 mL). Theaqueous layer was extracted with 25% propan-2-ol in DCM solution (3×30mL). The combined organic layers were dried using a hydrophobic frit andevaporated under reduced pressure to give the title compound (136.9 mg,0.360 mmol, 81% yield) as a pale yellow solid. LCMS (System B):t_(RET)=0.54 min, MH⁺=381.

Intermediate 158:5-(1-((1-acetylpyrrolidin-3-yl)methyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

tert-butyl3-((5-(hydroxymethyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-1-yl)methyl)pyrrolidine-1-carboxylate(for a preparation see Intermediate 159, 420 mg, 0.958 mmol) wasdissolved in DCM (7 mL) and 4M hydrochloric acid in 1,4-dioxane (0.958mL, 3.83 mmol) was added. The reaction mixture was stirred undernitrogen for 5 hours. The volatiles were removed by evaporation underreduced pressure and the sample was loaded in dichloromethane/methanolonto an SCX 10 g cartridge. Elutions of methanol, followed by 2M ammoniain methanol solution were used to purify the product. Appropriatefractions were combined, and concentrated to dryness. The product waspurified again on an SCX 10 g cartridge initially washed with methanol,followed by 33%, 50%, 66% and 100% 2M ammonia in methanol solutionelutions. The appropriate fractions were combined and evaporated underreduced pressure to give the crude product (309 mg, 0.913 mmol, 95%yield) as a pale yellow solid. The sample was dissolved in DMSO (3×1 mL)and purified by MDAP (Method B). The solvent was evaporated underreduced pressure to give the title compound (163 mg, 0.482 mmol, 50.3%yield) as a white solid. LCMS (System B): t_(RET)=0.48 min, MH⁺=339.

Intermediate 159: tert-butyl3-((5-(hydroxymethyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-1-yl)methyl)pyrrolidine-1-carboxylate

tert-butyl3-(((4-(hydroxymethyl)-2-nitrophenyl)amino)methyl)pyrrolidine-1-carboxylate(for a preparation see Intermediate 160, 1.0345 g, 2.94 mmol),5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparation seeintermediate 1, 0.404 g, 2.94 mmol) and sodium dithionite (1.538 g, 8.83mmol) were added to ethanol (8 mL) and water (4 mL). The reactionmixture was heated in a Biotage Initiator microwave using initial highabsorption setting to 100° C. for 5 hours. The reaction mixture waspartitioned between 25% propan-2-ol in DCM solution (30 mL) andsaturated sodium hydrogen carbonate solution (30 mL) and the layers wereseparated. The aqueous layer was extracted with 25% propan-2-ol in DCMsolution (3×30 mL). The organic layers were combined, dried using ahydrophobic frit and evaporated under reduced pressure. The sample wasloaded in dichloromethane and purified by Biotage SP4 SNAP 50 g silicausing a gradient of 2-12% dichloromethane-2M ammonia in methanol over 10column volumes followed by holding at 12% dichloromethane-2M ammonia inmethanol for 5 column volumes. The appropriate fractions were combinedand evaporated under reduced pressure to give the title compound (428.9mg, 0.978 mmol, 33.2% yield) as a colourless gum. LCMS (System B):t_(RET)=0.78 min, MH⁺=439.

Intermediate 160: tert-butyl3-(((4-(hydroxymethyl)-2-nitrophenyl)amino)methyl)pyrrolidine-1-carboxylate

(4-fluoro-3-nitrophenyl)methanol (521 mg, 3.04 mmol), tert-butyl3-(aminomethyl)pyrrolidine-1-carboxylate (850 mg, 4.24 mmol) and DIPEA(1.595 mL, 9.13 mmol) were added to tetrahydrofuran (THF) (3 mL) and thereaction mixture was heated in a Biotage Initiator microwave usinginitial high absorption setting to 120° C. for 3.5 hours. The reactionmixture was partitioned between DCM (25 mL) and saturated sodiumhydrogen carbonate solution (25 mL) and the layers were separated. Theaqueous layer was extracted with DCM (4×25 mL) and the combined organiclayers were dried using a hydrophobic frit and evaporated under reducedpressure. The sample was loaded in dichloromethane and purified bysilica gel column chromatography (50 g silica) using a gradient of 0-3%dichloromethane-2M ammonia in methanol over 10 column volumes. Theappropriate fractions were combined and evaporated under reducedpressure to give the title compound (1.0781 g, 3.07 mmol, 101% yield) asan orange oil. LCMS (System B): t_(RET)=1.03 min, MH⁻=350.

Intermediate 161:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazole-5-carbaldehyde

A round bottom flask was charged with5-(5-(hydroxymethyl)-1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 162, 360 mg, 1.019 mmol), DCM (20mL) and Dess-Martin periodinane (432 mg, 1.019 mmol). The mixture wasstirred at room temperature overnight. The mixture was diluted with DCMand saturated sodium bicarbonate added before the layers were mixed andseparated. The organics were washed with brine before being passedthrough a hydrophobic frit and concentrated in vacuo to give anoff-white solid. The sample was loaded in dichloromethane and purifiedby silica gel column chromatography (25 g silica) using a gradient of0-100% ethyl acetate-cyclohexane over 15 CV followed by 0-10%dichloromethane-methanol for 15 CV. The appropriate fractions werecombined and evaporated in vacuo to give the title compound (345 mg,0.884 mmol, 87% yield) as a white solid. LCMS (System A): t_(RET)=0.65min, MH⁺=352.

Intermediate 162:5-(5-(hydroxymethyl)-1-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

A round bottom flask was charged with1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, 574 mg, 3.23 mmol), sodium hydrosulfite (1.8 g,10.34 mmol), water (10 mL), and a solution of(3-nitro-4-((tetrahydro-2H-pyran-4-yl)amino)phenyl)methanol (for apreparation see intermediate 163, 740 mg, 2.93 mmol) in ethanol (20 mL).The vessel was fitted with an air condenser and the slurry heated at100° C. overnight. The mixture was diluted with water and EtOAc, thelayers mixed and separated before the organic layer was passed through ahydrophobic frit and concentrated in vacuo to give a yellow oil. Thesample was loaded in dichloromethane and purified by silica gel columnchromatography (25 g silica) using a gradient of 0-100% ethylacetate-cyclohexane over 15 CV followed by 0-10% 2M ammonia inmethanol-dichloromethane for 15CV. The appropriate fractions werecombined and evaporated in vacuo to give the title compound (360 mg,1.019 mmol, 34.7% yield) as a colourless oil. LCMS (System A):t_(RET)=0.47 min, MH⁺=354.

Intermediate 163:(3-nitro-4-((tetrahydro-2H-pyran-4-yl)amino)phenyl)methanol

A round bottom flask was charged with (4-fluoro-3-nitrophenyl)methanol(1 g, 5.84 mmol), tetrahydro-2H-pyran-4-amine, hydrochloride (0.9 g,6.54 mmol, J&W PharmLab), DMF (10 mL) and DIPEA (4.1 mL, 23.48 mmol). Anair condenser was fitted and the slurry warmed to 70° C. overnight. Themixture was cooled to room temperature, diluted with EtOAc and theorganics washed with 1M HCl followed by brine before being passedthrough a hydrophobic frit. The filtrate was concentrated in vacuo togive an orange oil. The sample was loaded in dichloromethane andpurified by silica gel column chromatography (50 g silica) using agradient of 0-100% ethyl acetate-cyclohexane over 15 CV. The appropriatefractions were combined and evaporated in vacuo to give an orange oil.The sample was dissolved in EtOAc before being washed with 10% LiClsolution. the organics were passed through a hydrophobic frit beforebeing concentrated in vacuo to give the title compound (740 mg, 2.93mmol, 50.2% yield) as an orange solid. LCMS (System A): t_(RET)=0.71min, MH⁺=253.

Intermediate 164:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

5-(5-(hydroxymethyl)-1-((4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 165, 506 mg, 1.323 mmol) wasdissolved in dichloromethane (DCM) (5 mL) and Dess-Martin periodinane(561 mg, 1.323 mmol) added. This mixture was stirred under nitrogen for18 hrs. The mixture was then partitioned between ethyl acetate (100 mL)and saturated sodium bicarbonate (100 mL) and the phases separated. Theaqueous phase was then extracted twice with ethyl acetate (100 mL×2) andthe organics combined. Organics were then washed with brine (100 mL) anddried using a hydrophobic frit. Solvent was then removed in vacuoyielding the title compound (457 mg, 1.201 mmol, 91% yield) as anoff-white solid. LCMS (System B): t_(RET)=0.68 min, MH⁺=381.

Intermediate 165:5-(5-(hydroxymethyl)-1-((4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(4-(((4-methylmorpholin-2-yl)methyl)amino)-3-nitrophenyl)methanol (for apreparation see Intermediate 166, 1.6 g, 5.69 mmol),1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, 0.860 g, 5.69 mmol) and sodium dithionite (2.97 g,17.06 mmol) was added to a microwave vial along with ethanol (8 mL) andwater (8 mL) and heated to 100° C. for 5 hrs. The sample was thenpartitioned between ethyl acetate (75 mL) and saturated sodium carbonate(75 mL) and the phases separated. The organic phases was retained andaqueous phase extracted twice with ethyl acetate (75 mL×2). The organicswere then combined, dried using a hydrophobic frit then solvents removedin vacuo. This yielded a pale yellow solid. This was dissolved indichloromethane. The sample was loaded onto a Biotage SNAP 100 g Silicacartridge and eluted using a gradient of 0%-10% methanolic ammonia/DCMfollowed by 10%-15% methanolic ammonia/DCM. The appropriate fractionswere then combined yielding the title compound (506 mg, 1.323 mmol,23.26% yield) as a yellow oil. LCMS (System B): t_(RET)=0.61 min,MH⁺=383.

Intermediate 166:(4-(((4-methylmorpholin-2-yl)methyl)amino)-3-nitrophenyl)methanol

(4-fluoro-3-nitrophenyl)methanol (1.041 g, 6.08 mmol) and(4-methylmorpholin-2-yl)methanamine (2.376 g, 18.25 mmol) were dissolvedin THF (5 mL) and treated with N,N-diisopropylethylamine (3.19 mL, 18.25mmol). This mixture was heated in a microwave reactor at 120° C. for 1hr. The dark orange mixture was then partitioned between ethyl acetate(75 mL) and saturated sodium bicarbonate (75 mL). The phases were thenseparated and the aqueous phase extracted twice with ethyl acetate (75mL×2). The organics were then combined and dried using a hydrophobicfrit then solvents removed in vacuo. This yielded the title compound(1.6 g, 5.69 mmol, 93% yield) as an orange solid. LCMS (System B):t_(RET)=0.69 min, MH⁺=282.

Intermediate 167:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((4-methylmorpholin-3-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

5-(5-(hydroxymethyl)-1-((4-methylmorpholin-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 168, 473 mg, 1.237 mmol) wasdissolved in DCM and Dess-Martin periodinane (525 mg, 1.237 mmol) added.This mixture was stirred at room temperature for 20 hrs. Saturatedsodium bicarbonate (100 mL) was then added and the mixture stirred for15 min. To the mixture was added ethyl acetate (100 mL) and the phasesseparated. The aqueous phase was then extracted twice with ethyl acetate(100 mL×2). The organics were then combined, dried using a hydrophobicfrit then solvent removed in vacuo; to yield the crude title compound(450 mg, 1.18 mmol, 96% yield) as a yellow solid which was used withoutfurther purification. LCMS (System B): t_(RET)=0.65 min, MH⁺=381.

Intermediate 168:5-(5-(hydroxymethyl)-1-((4-methylmorpholin-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(4-(((4-methylmorpholin-3-yl)methyl)amino)-3-nitrophenyl)methanol (for apreparation see Intermediate 169, 2.83 g, 10.06 mmol) and1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, 1.977 g, 13.08 mmol) were dissolved in ethanol (16mL) along with sodium dithionite (5.25 g, 30.2 mmol) and water (16 mL).The mixtures were heated in a microwave reactor for 5 hrs at 120° C. Theyellow mixture was then partitioned between ethyl acetate (150 mL) andsaturated sodium bicarbonate (150 mL) and the phases separated. Theaqueous phase was extracted twice with ethyl acetate (150 mL×2) and theorganics combined. The organics were then dried using a hydrophobicfrit. The solvent was then removed in vacuo yielding a yellow oil. Theyellow oil was dissolved in dichloromethane and loaded onto a biotageSNAP 100 g silica column and eluted using a gradient of 0%-15% 2Mmethanolic ammonia/DCM. Appropriate fractions were combined and solventsremoved in vacuo to yield the title compound (473 mg, 1.24 mmol, 12.3%yield) as a yellow oil. LCMS (System B): t_(RET)=0.58 min, MH⁺=383.

Intermediate 169:(4-(((4-methylmorpholin-3-yl)methyl)amino)-3-nitrophenyl)methanol

(4-fluoro-3-nitrophenyl)methanol (1.77 g, 10.34 mmol, Aldrich) and(4-methylmorpholin-3-yl)methanamine (4.14 mL, 31.0 mmol, Chess FineOrganics) were dissolved in tetrahydrofuran (THF) (5 mL) and treatedwith N,N-diisopropylethylamine (5.42 mL, 31.0 mmol). This mixture washeated in a microwave reactor at 120° C. for 1 hr. The dark orangemixture was then partitioned between ethyl acetate (150 mL) andsaturated sodium bicarbonate (150 mL). The phases were then separatedand the aqueous phase extracted twice with ethyl acetate (150 mL×2). Theorganics were then combined and dried using a hydrophobic frit thensolvents removed in vacuo. This yielded the title compound(4-(((4-methylmorpholin-3-yl)methyl)amino)-3-nitrophenyl)methanol (2.83g, 10.06 mmol, 97% yield) as a dark red oil. LCMS (System B):t_(RET)=0.70 min, MH⁺=282.

Intermediate 170:(R)-5-(6-(hydroxymethyl)-1-(piperidin-3-ylmethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

HCl 5 M in IPA (9 ml, 45.0 mmol) was added to (S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-(hydroxymethyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate(525 mg, 0.979 mmol, for a preparation see Intermediate 100) and theresulting solution was stirred at rt for 2 hr. After this time thesolution was evaporated under reduced pressure to obtain a palepink-white solid. The solid was dissolved in MeOH and loaded into anSCX-2 cartridge (20 g). The cartridge was flushed with MeOH (3×CV)followed by 2 M Ammonia in MeOH (3×CV). The basic fractions werecombined to obtain the title compound N31482-98-1, 287 mg, 80% yield asa pale yellow oil. LCMS (System B): t_(RET)=0.59 min, MH⁺=367.

Intermediate 171: (3-((cyclopropylmethyl)amino)-4-nitrophenyl)methanol

(3-fluoro-4-nitrophenyl)methanol (1 g, 5.84 mmol) was dissolved in2-methyl tetrahydrofuran (10 mL) and DIPEA (3.06 mL, 17.52 mmol) andcyclopropylmethylamine (0.76 mL, 8.76 mmol) were added. The resultingmixture was stirred at 80° C. overnight.

The reaction mixture was partitioned between DCM and saturated aqueoussodium hydrogen carbonate solution. The aqueous layer was extracted withDCM. The combined organics were washed with saturated aqueous sodiumhydrogen carbonate solution, dried using a hydrophobic frit andevaporated in vacuo to give an orange oil which solidified on standing(1.65 g). The crude product was purified by chromatography on silica(100 g) using a 0-50% ethyl acetate/cyclohexane gradient. Appropriatefractions were combined and evaporated to give the title compound (1.28g) as a bright orange solid. LCMS (System A): t_(RET)=0.98 min; MH⁺=223.

Intermediate 172:5-(1-(cyclopropylmethyl)-6-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(3-((cyclopropylmethyl)amino)-4-nitrophenyl)methanol (for a preparationsee Intermediate 171, 1.28 g, 5.76 mmol) and1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2 N31961-84-1, 1.13 g, 7.49 mmol) were dissolved inethanol (20 mL) and water (10 mL). Sodium dithionate (3.56 g, 17.28mmol) was added and the reaction was heated at 80° C. for 2 hours. Thereaction mixture was partitioned between ethyl acetate and saturatedaqueous sodium bicarbonate solution. The organic layer was washed withsaturated aqueous sodium bicarbonate solution, dried using a hydrophobicfrit and evaporated to give the title compound (1.39 g) a beige solid.LCMS (System A): t_(RET)=0.49 min; MH⁺=324.

Intermediate 173:1-(cyclopropylmethyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-6-carbaldehyde

5-(1-(cyclopropylmethyl)-6-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 172, 1.38 g, 4.27 mmol) wasdissolved in chloroform (50 mL) and manganese dioxide (3.71 g, 42.7mmol) was added. The resulting suspension was stirred at roomtemperature under nitrogen overnight. The reaction mixture was filteredthrough celite and evaporated to give the title compound (1.38 g) as apale yellow oil, which solidified on standing. LCMS (System A):t_(RET)=0.76 min; MH⁺=322.

Intermediate 174:(4-nitro-3-(((tetrahydro-2H-pyran-3-yl)methyl)amino)phenyl)methanol

(tetrahydro-2H-pyran-3-yl)methanamine hydrochloride (1.33 g, 8.77 mmol,Enamine) was added to a mixture of (3-fluoro-4-nitrophenyl)methanol (1g, 5.84 mmol, Apollo) and DIPEA (4.08 mL, 23.36 mmol) in 2-methyltetrahydrofuran (10 mL). The resulting mixture was stirred at 80° C.overnight. The reaction mixture was partitioned between DCM andsaturated aqueous sodium hydrogen carbonate solution. The aqueous layerwas extracted with DCM. The combined organics were washed with saturatedaqueous sodium hydrogen carbonate solution, dried using a hydrophobicfrit and evaporated in vacuo to give an orange oil (1.74 g). The crudeproduct was purified by chromatography on silica (100 g) using a 0-100%ethyl acetate/cyclohexane gradient. Appropriate fractions were combinedand evaporated to give the title compound (1.35 g) as an orange oil.LCMS (System A): t_(RET)=0.88 min; MH⁺=267.

Intermediate 175 and 176:5-(6-(hydroxymethyl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(4-nitro-3-(((tetrahydro-2H-pyran-3-yl)methyl)amino)phenyl)methanol (fora preparation see Intermediate 174, 134 g, 5.03 mmol) and1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, 0.99 g, 6.54 mmol) were dissolved in ethanol (20 mL)and water (10 mL). Sodium dithionate (3.11, 15.09 mmol) was added andthe reaction was heated at 80° C. for 2 hours. The reaction mixture waspartitioned between ethyl acetate and sat. aqueous sodium bicarbonatesolution. The organic layer was washed with sat. aqueous sodiumbicarbonate solution, dried using a hydrophobic frit and evaporated togive a beige solid (1.01 g). The crude product was purified bychromatography on silica (100 g) using a 0-50% (20% ammonia methanol indichloromethane)/dichloromethane gradient. Appropriate fractions werecombined and evaporated to give a colourless oil (0.99 g) whichsolidified on standing. LCMS (System A): t_(RET)=0.47 min; MH+=328. Thismaterial was separated into its two component enantiomers by preparativechiral HPLC. The racemate was dissolved in ethanol and purified bychiral chromatography (stationary phase: Chiralpak AD-H (250×30 mm, 5micron), mobile phase: hexane/ethanol (+0.2% v/v isopropylamine)).Appropriate fractions were combined and evaporated to give the twoenantiomers: Intermediate 175: 426 mg, yellow solid. LCMS (System A):t_(RET)=0.47 mins; MH+=368. Intermediate 176: 488 mg, yellow solid. LCMS(System A): t_(RET)=0.47 mins; MH+=368. Absolute stereochemistry was notassigned.

Intermediate 177a:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazole-6-carbaldehyde(Isomer 1)

5-(6-(hydroxymethyl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 175 [isomer 1], 423 mg, 1.151 mmol)was dissolved in chloroform (10 mL) and manganese dioxide (1 g, 11.50mmol) was added. The resulting suspension was stirred at roomtemperature under nitrogen for 6 hours. The reaction mixture wasfiltered through celite and evaporated to give the title compound (400mg) as a pale yellow solid. LCMS (System A): t_(RET)=0.73 mins; MH+=366.

Intermediate 177b:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazole-6-carbaldehyde(Isomer 2)

5-(6-(hydroxymethyl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 176, [Isomer 2], 484 mg, 1.317 mmol)was dissolved in chloroform (10 mL) and manganese dioxide (1.14 g, 13.11mmol) was added. The resulting suspension was stirred at roomtemperature under nitrogen overnight. LCMS showed ˜15% starting materialremained. A further portion of manganese dioxide (0.6 g, 6.90 mmol) wasadded and the reaction was stirred at room temperature, under nitrogen,for 5 hours. The reaction mixture was filtered through celite andevaporated to give the title compound (422 mg) as a yellow solid. LCMS(System A): t_(RET)=0.73 mins; MH+=366.

Intermediate 178: 1-(tetrahydro-2H-pyran-4-yl)ethanamine, Hydrochloride

A black suspension ofN-(4-methoxybenzyl)-1-(tetrahydro-2H-pyran-4-yl)ethanamine (for apreparation see Intermediate 179, 4.95 g, 18.86 mmol) and 10% w/wpalladium on carbon (0.401 g, 1.886 mmol) in ethanol (100 mL) wasstirred under hydrogen for 2 days. The reaction mixture was passedthrough celite column, rinsed with EtOH, and 2M aq. HCl (10 mL) added.The resulting solution was stirred for 20 min, and evaporated in vacuoto afford the title compound as a white solid. The total yield of thereaction was 80%. 8.13 (br.s, 8.13, 3H), 3.88 (dd, J=3.5, 11.0 Hz, 2H),3.24 (tdd, 2.0, 4.0, 12.0 Hz, 2H), 3.06-2.92 (m, 1H), 1.79-1.66 (m, 1H),1.66-1.52 (m, 2H), 1.34-1.19 (m, 2H), 1.16 (d, J=6.5 Hz, 3H).

Intermediate 179:N-(4-methoxybenzyl)-1-(tetrahydro-2H-pyran-4-yl)ethanamine

To a stirred solution of 1-(tetrahydro-2H-pyran-4-yl)ethanone (3.2 g,24.97 mmol) in DCM) (50 mL was added (4-methoxyphenyl)methanamine (6.87g, 50.1 mmol). The resulting yellow solution was stirred for 4.5 h andsodium triacetoxyborohydride (10 g, 48.6 mmol) added. The whitesuspension was stirred. The reaction mixture was partitioned between DCMand aq. sat. NaHCO₃. The organic layer was removed and the aqueousextracted 3 times with DCM. The combined organic phases were passedthrough a hydrophobic frit and concentrated in vacuo to give a yellowoil. The oil was dissolved in DCM, purified by silica gel chromatographyeluting with EtOAc:EtOH (7.5-25%) and evaporated in vacuo to give thetitle compound as a yellow oil. The total yield of the reaction was 80%.LCMS (System A): t_(RET)=1.27 min, MH⁺=366.

Intermediate 180:3-((1,3-dimethoxypropan-2-yl)amino)-4-nitrophenyl)methanol

(1,4-dioxan-2-yl)methanamine (3.08 g, 26.3 mmol) was added to a mixtureof (3-fluoro-4-nitrophenyl)methanol (4.1 g, 24 mmol), DIPEA (9.18 mL,52.6 mmol) in THF (30 mL). The resulting mixture was stirred at 80° C.overnight. The reaction mixture was partitioned between DCM (100 mL) andsaturated aqueous sodium hydrogen carbonate solution (100 mL) and thelayers separated. The aqueous layer was extracted with DCM (3×100 mL)and the combined organic layers were passed through a phase separatorand evaporated in vacuo to afford ˜10 g of a crude mixture of the titlecompound. Used at this purity in next step. LCMS (System A):t_(RET)=0.88 min, MH⁺=271.

Intermediate 181: (S)-cyclopentyl4-methyl-2-((4-((1-methylpiperidin-4-yl)amino)-3-nitrobenzyl)amino)pentanoate

To a solution of (S)-cyclopentyl2-((4-fluoro-3-nitrobenzyl)amino)-4-methylpentanoate (For a preparationsee Intermediate 144, 500 mg, 1.419 mmol) in THF (9 mL) was added1-methylpiperidin-4-amine (486 mg, 4.26 mmol) and DIPEA (0.743 mL, 4.26mmol), and the reaction mixture heated in a microwave to 120° C. for atotal of 90 mins. Further 1-methylpiperidin-4-amine (486 mg, 4.26 mmol)was added and the reaction mixture heated for a further 30 mins at 120°C. The reaction mixture was partitioned between DCM (2×100 mL) andsaturated aqueous sodium bicarbonate solution (100 mL). The organiclayers were combined, dried using a hydrophobic frit and evaporatedunder reduced pressure. The sample was loaded in dichloromethane andpurified by silica gel column chromatography using a gradient of 0-5%DCM-2M ammonia in methanol over 10 column volumes followed by holding at5% DCM-2M ammonia in methanol for 5 column volumes. The appropriatefractions were combined and evaporated under reduced pressure to givethe title compound (456 mg) as an orange gum. LCMS (System A):t_(RET)=0.70 min, MH⁺=447.

The following Intermediates were prepared in a similar manner toIntermediate 181 using the appropriate commercially available amine, andthe appropriate fluorophenyl intermediate as shown in the table below:

Intermediate 182: (S)-cyclopentyl4-methyl-2-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)benzyl)amino)pentanoate

(prepared from: Intermediate 144) System A, 0.98 min, MH⁺=448; Yield:603 mg, 85%

Intermediate 183: (2S)-cyclopentyl4-methyl-2-((4-(((4-methylmorpholin-2-yl)methyl)amino)-3-nitrobenzyl)amino)pentanoate

(prepared from: Intermediate 144) System B, 1.37 min, MH⁺=463; Yield:1.250 g, 98%

Intermediate 184: (S)-cyclopentyl2-((4-((2-methoxyethyl)amino)-3-nitrobenzyl)amino)-4-methylpentanoate

(prepared from: Intermediate 144) System A, 0.94 min, MH⁺=408; Yield:353 mg, 61%

Intermediate 185: (S)-cyclopentyl2-((4-((2-(dimethylamino)ethyl)amino)-3-nitrobenzyl)amino)-4-methylpentanoate

(prepared from: Intermediate 144) System A, 0.69 min, MH⁺=421 Yield: 543mg, 91%

Intermediate 186: (S)-cyclopentyl2-((4-((3-hydroxypropyl)amino)-3-nitrobenzyl)amino)-4-methylpentanoate

(prepared from: Intermediate 144) System A, 0.85 min, MH⁺=408; Yield:445 mg, 77%

Intermediate 187: (S)-cyclopentyl4-methyl-2-((4-(methylamino)-3-nitrobenzyl)amino)pentanoate

(prepared from: Intermediate 144) System A, 0.89 min, MH⁺=354; Yield:171 mg, 40%

Intermediate 188: (S)-Tert-butyl2-((4-fluoro-3-nitrobenzyl)amino)-4-methylpentanoate

To a solution of 4-fluoro-3-nitrobenzaldehyde (1 g, 5.91 mmol) and(S)-tert-butyl 2-amino-4-methylpentanoate hydrochloride (1.46 g, 6.50mmol) in DCM (25 mL) was added acetic acid (1.016 mL, 17.74 mmol) andthe reaction mixture stirred under nitrogen for 1 hour. Sodiumtriacetoxyborohydride (2.507 g, 11.83 mmol) was added portion-wise, andthe reaction mixture stirred at room temperature overnight. Saturatedaqueous sodium bicarbonate solution (50 mL) was added slowly, andstirring continued until fizzing had stopped. The resulting suspensionwas extracted with DCM (3×50 mL). The organic layers were combined,dried using a hydrophobic frit and evaporated under reduced pressure.The sample was loaded in DCM and purified by SPE (silica, 100 g) using agradient of 0-50% EtOAc in cyclohexane. The appropriate fractions werecombined and evaporated under reduced pressure to give the titlecompound (1.66 g, 4.88 mmol) as a yellow gum. LCMS (System A):t_(RET)=0.95 min, MH⁺=341.

Intermediate 189: (S)-tert-Butyl4-methyl-2-((4-((1-methylpiperidin-4-yl)amino)-3-nitrobenzyl)amino)pentanoate

To a solution of (S)-tert-butyl2-((4-fluoro-3-nitrobenzyl)amino)-4-methylpentanoate (For an examplepreparation see Intermediate 188, 250 mg, 0.734 mmol) in THF (3.5 mL)was added 1-methylpiperidin-4-amine (252 mg, 2.203 mmol) and DIPEA(0.385 mL, 2.203 mmol), and the reaction mixture heated under microwaveconditions to 120° C. for 30 min. The reaction mixture was partitionedbetween DCM (2×20 mL) and saturated aqueous sodium bicarbonate solution(20 mL). The organic layers were combined, dried using a hydrophobicfrit and blown down under a stream of nitrogen. The sample was loaded inDCM and purified by SPE (silica, 25 g) using a gradient of 0-10% (2 Mammonia in MeOH) in DCM. The appropriate fractions were combined andevaporated under reduced pressure to give the title compound (127 mg,0.292 mmol). LCMS (System B): t_(RET)=1.09 min, MH⁺=435.

Intermediate 190: (S)-Cyclopentyl4-methyl-2-((4-nitro-3-(((tetrahydro-2H-pyran-4-yl)methyl)amino)benzyl)amino)pentanoate

A solution of 3-fluoro-4-nitrobenzaldehyde (500 mg, 2.96 mmol) and(S)-cyclopentyl 2-amino-4-methylpentanoate 4-methylbenzenesulfonate (Foran example preparation see Intermediate 3, 1208 mg, 3.25 mmol) in DCM(20 mL) was stirred under nitrogen for 1.5 hours. Sodiumtriacetoxyborohydride (1253 mg, 5.91 mmol) was added portionwise, andthe reaction mixture stirred at room temperature overnight. Saturatedaqueous sodium bicarbonate solution (50 mL) was added slowly, and thereaction stirring continued until fizzing had stopped. The resultingsuspension was extracted with DCM (3×50 mL). The organic layers werecombined, dried using a hydrophobic frit and evaporated under reducedpressure. The sample was loaded in dichloromethane and purified by SPE(silica, 50 g) using a gradient of 0-50% EtOAc in cyclohexane. Theappropriate fractions were combined and blown down under a stream ofnitrogen to give a yellow gum. To this material in THF (4 mL) was added(tetrahydro-2H-pyran-4-yl)methanamine (260 mg, 2.257 mmol) and DIPEA(0.395 mL, 2.259 mmol), and the reaction mixture heated under microwaveconditions at 120° C. for 30 min. The reaction mixture was partitionedbetween dichloromethane (3×50 mL) and saturated aqueous sodiumbicarbonate solution (50 mL). The organic layers were combined, driedusing a hydrophobic frit and blown down under a stream of nitrogen. Thesample was loaded in dichloromethane and purified by SPE (silica, 50 g)using a gradient of 0-5% (2 M ammonia in MeOH) in DCM. The appropriatefractions were combined and blown down under a stream of nitrogen togive the crude product. The crude product was again partitioned betweendichloromethane (3×50 mL) and saturated aqueous sodium bicarbonatesolution (50 mL). The organic layers were combined, dried using ahydrophobic frit and blown down under a stream of nitrogen to give thetitle compound (522 mg, 1.166 mmol). LCMS (System B): t_(RET)=1.49 min,MH⁺=448

Intermediate 191: (S)-Neopentyl2-(((benzyloxy)carbonyl)amino)-3-hydroxypropanoate

To a solution of (S)-2-(((benzyloxy)carbonyl)amino)-3-hydroxypropanoicacid (2.27 g, 9.49 mmol), EDC (2.18 g, 11.39 mmol) and HOBT (1.74 g,11.39 mmol) in DMF (20 mL) was added DIPEA (3.31 mL, 18.98 mmol) and2,2-dimethylpropan-1-ol (8.36 g, 95 mmol), and the reaction mixturestirred under nitrogen at room temperature overnight. The reactionmixture was partitioned between ethyl acetate (50 mL) and saturatedaqueous sodium bicarbonate solution (50 mL). The organic layer was thenwashed with 1 M aqueous hydrochloric acid (50 mL), water (50 mL) andbrine (50 mL). The organic layer was then dried using a hydrophobic fritand blown down under a stream of nitrogen. The sample was loaded indichloromethane and purified by SPE (silica, 100 g) using a gradient of0-50% EtOAc in cyclohexane. The appropriate fractions were combined andevaporated under reduced pressure to give the title compound (951 mg,3.07 mmol) as a white solid. LCMS (System B): t_(RET)=1.06 min, MH⁺=310.

Intermediate 192: (S)-Neopentyl 2-amino-3-methoxypropanoate

To a mixture of (S)-neopentyl2-(((benzyloxy)carbonyl)amino)-3-hydroxypropanoate (For an examplepreparation see Intermediate 191, 947 mg, 3.06 mmol) and silver oxide(1064 mg, 4.59 mmol) in dry acetonitrile (20 mL) was added methyl iodide(1.914 mL, 30.6 mmol), and the reaction mixture heated at 90° C. undernitrogen for two nights. The reaction mixture was allowed to cool toroom temperature, and the solid removed by filtration. The resultingsolution was evaporated under reduced pressure. The sample was loaded indichloromethane and purified by SPE (silica, 50 g) using a gradient of0-25% EtOAc in cyclohexane. The appropriate fractions were combined andevaporated under reduced pressure. A solution of this material (347 mg)in ethanol (10 mL) was hydrogenated using an H-cube (settings: 20° C., 1bar, 1 mL/min flow rate) and 10% Pd/C CatCart 30 as the catalyst. Thereaction mixture was blown down under a stream of nitrogen and dried ina vacuum oven to give the title compound (199 mg, 1.052 mmol). LCMS(System B): t_(RET)=0.81 min, MH⁺=190 no UV chromophore.

Intermediate 193: (3-Nitro-4-((oxetan-3-ylmethyl)amino)phenyl)methanol

A round bottom flask was charged with (4-fluoro-3-nitrophenyl)methanol(770 mg, 4.50 mmol), oxetan-3-ylmethanamine hydrochloride (753 mg, 6.09mmol), THF (10 mL) and DIPEA (2.3 mL, 13.17 mmol). An air condenser wasfitted and the slurry warmed to 62° C. overnight. The mixture was cooledto room temperature before DMF (2 mL) was added. The mixture was warmedto 70° C. for 4 days. The mixture was cooled to room temperature,diluted with EtOAc and the organics washed with water followed by brinebefore being passed through a hydrophobic frit. The filtrate wasconcentrated in vacuo to give an orange oil. The sample was loaded indichloromethane and purified by SPE (silica, 25 g) using a gradient of0-100% EtOAc in cyclohexane. The appropriate fractions were combined andevaporated in vacuo to give the title compound (584 mg, 2.451 mmol) asan orange solid. LCMS (System A): t_(RET)=0.61 min, MH⁺=239.

Intermediate 194:5-(5-(Hydroxymethyl)-1-(oxetan-3-ylmethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

A microwave vial was charged with5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (For an examplepreparation see Intermediate 1, 504 mg, 3.68 mmol), sodium hydrosulfite(1.5 g, 8.62 mmol), water (5.00 mL), and a solution of(3-nitro-4-((oxetan-3-ylmethyl)amino)phenyl)methanol (For an examplepreparation see Intermediate 193, 584 mg, 2.451 mmol) in ethanol (10mL). The vial was capped and the slurry irradiated at 100° C. for 5hours. The mixture was diluted with water and chloroform/IPA (3:1), thelayers mixed and separated before the organic layer was passed through ahydrophobic frit and concentrated in vacuo to give a yellow oil. Thesample was loaded in dichloromethane and purified by SPE (silica, 25 g)using a gradient of 0-100% EtOAc in cyclohexane. The appropriatefractions were combined and evaporated in vacuo to give the titlecompound (178 mg, 0.547 mmol) as a colourless oil which solidified. LCMS(System A): t_(RET)=0.40 min, MH⁺=326.

Intermediate 195:2-(5-Methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(oxetan-3-ylmethyl)-1H-benzo[d]imidazole-5-carbaldehyde

To a solution of5-(5-(hydroxymethyl)-1-(oxetan-3-ylmethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(For an example preparation see Intermediate 194,_180 mg, 0.553 mmol) inDCM (15 mL) was added 45% iodoxybenzoic acid (stabilised by benzoic acidand isophthalic acid) (379 mg, 0.609 mmol), portionwise. The reactionmixture was stirred at room temperature for 4 nights. The reactionmixture was partitioned between 10% isopropanol in dichloromethane (100mL) and saturated aqueous sodium bicarbonate solution (3×100 mL). Theorganic layer was passed through a hydrophobic frit and blown down undera stream of nitrogen to give the title compound (152 mg, 0.470 mmol) asa pale yellow gum. LCMS (System B): t_(RET)=0.60 min, MH⁺=324.

Intermediate 196: (S)-Cyclopentyl 2-amino-3-methoxypropanoate

(S)-2-amino-3-methoxypropanoic acid hydrochloride (3.3131 g, 21.30 mmol)was added to cyclopentanol (30 mL) and the suspension was brought to −5°C. using a dry ice/acetone bath. After stirring at this temperature forten minutes, thionyl chloride (3.57 mL, 49.0 mmol) was added dropwise.The suspension was left stirring and allowed to warm up to roomtemperature. The reaction mixture was warmed to 60° C. and stirred atthis temperature for 24 hours. Volatiles were removed from the reactionmixture under reduced pressure. Hot EtOAc was added with the intentionof performing a recrystallisation. The material did not dissolve so thesuspension was filtered off, washed on the filter, and dried in a vacuumoven to give the title compound (4.49 g, 0.24 mmol) as a white solid. ¹HNMR (d₆-DMSO, 293 K): δ 1.51-1.76 (m, 6H) 1.76-1.92 (m, 2H) 3.29 (s, 3H)3.75 (d, J=3.4 Hz, 2H) 4.21 (t, J=3.4 Hz, 1H) 5.17-5.21 (m, 1H) 8.64(br.s., 3H).

Intermediate 197:5-(1-Ethyl-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

To a mixture of 1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde(For an example preparation see Intermediate 2, 1 g, 6.62 mmol) andsodium hydrosulfite (3.46 g, 19.85 mmol) was added a solution of(4-(ethylamino)-3-nitrophenyl)methanol (For an example preparation seeIntermediate 70, 1.298 g, 6.62 mmol) in ethanol (30 mL), followed bywater (15 mL). The reaction mixture was heated at 80° C. overnight. Thereaction mixture was partitioned between saturated aqueous sodiumhydrogen carbonate solution (150 mL) and 3:1 chloroform:isopropanol(3×150 mL). The organic layers were combined, dried using a hydrophobicfrit and evaporated under reduced pressure. The sample was loaded indichloromethane and purified by SPE (silica, 100 g) silica using agradient of 0-12% (2 M ammonia in methanol) in DCM. The appropriatefractions were combined and evaporated under reduced pressure to givethe title compound (1.05 g, 3.53 mmol) as an off-white foam. LCMS(System B): t_(RET)=0.64 min, MH⁺=298

Intermediate 198:2-(1,5-Dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-ethyl-1H-benzo[d]imidazole-5-carbaldehyde

To a solution of5-(1-ethyl-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(For an example preparation see Intermediate 197, 1.05 g, 3.53 mmol) inDCM (50 mL) was added 45% iodoxybenzoic acid (stabilised by benzoic acidand isophthalic acid) (2.417 g, 3.88 mmol), portionwise. The reactionmixture was stirred at room temperature for 4 nights. The reactionmixture was partitioned between 10% isopropanol in dichloromethane (250mL) and saturated aqueous sodium bicarbonate solution (3×200 mL). Theorganic layer was dried using a hydrophobic frit and blown down under astream of nitrogen to give the title compound (856 mg, 2.90 mmol) as apale yellow solid. LCMS (System B): t_(RET)=0.72 min, MH⁺=296.

Intermediate 199: (4-((2-Methoxyethyl)amino)-3-nitrophenyl)methanol

A round bottom flask was charged with (4-fluoro-3-nitrophenyl)methanol(3.2 g, 18.70 mmol), 2-methoxyethanamine (2.107 g, 28.0 mmol), THF (30mL) and DIPEA (9.80 mL, 56.1 mmol). An air condenser was fitted and theslurry warmed to 62° C. overnight. The mixture was cooled to roomtemperature and diluted with EtOAc. The organics were washed with waterbefore being passed through a hydrophobic frit and concentrated in vacuoto give an orange oil. The sample was loaded in dichloromethane andpurified by SPE (silica, 100 g) using a gradient of 0-80% EtOAc incyclohexane. The appropriate fractions were combined and evaporated invacuo to give the title compound (3.48 g, 15.38 mmol) as a orange oilwhich solidified. LCMS (System A): t_(RET)=0.70 min, MH⁺=227.

Intermediate 200:5-(5-(Hydroxymethyl)-1-(2-methoxyethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

A round bottom flask was charged with5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (For an examplepreparation see Intermediate 1, 2.53 g, 18.46 mmol), sodium hydrosulfite(9.37 g, 53.8 mmol), water (25.00 mL), and a solution of(4-((2-methoxyethyl)amino)-3-nitrophenyl)methanol (For an examplepreparation see Intermediate 199, 3.48 g, 15.38 mmol) in ethanol (50mL). The vessel was fitted with an air condenser and warmed to 100° C.overnight. The mixture was diluted with water and chloroform/IPA (3:1),the layers mixed and separated before the organic layer was passedthrough a hydrophobic frit and concentrated in vacuo to give a whitesolid. The sample was loaded in dichloromethane and purified by SPE(silica, 25 g) using a gradient of 0-100% EtOAc in cyclohexane. Theappropriate fractions were combined and evaporated in vacuo to give thetitle compound (3.58 g, 11.42 mmol) as a colourless oil whichsolidified. LCMS (System A): t_(RET)=0.44 min, MH⁺=314.

Intermediate 201:1-(2-Methoxyethyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde

To a solution of5-(5-(hydroxymethyl)-1-(2-methoxyethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(For and example preparation see Intermediate 200, 3.38 g, 10.79 mmol)in DCM (150 mL) was added 45% iodoxybenzoic acid (stabilised by benzoicacid and isophthalic acid) (7.38 g, 11.87 mmol), portionwise. Thereaction mixture was stirred at room temperature for 4 nights. Thereaction mixture was partitioned between DCM (200 mL) and saturatedaqueous sodium bicarbonate solution (200 mL). The aqueous layercontained a large amount of solid, so was further extracted with 10%isopropanol in dichloromethane (2×200 mL). The organic layers werecombined, washed with aqueous sodium bicarbonate solution (2×200 mL),dried using a hydrophobic frit and evaporated under reduced pressure togive the title compound (2.7 g, 8.67 mmol) as a pale brown solid. LCMS(System B): t_(RET)=0.66 min, MH⁺=312.

Intermediate 202:5-(5-(Hydroxymethyl)-1-(2-methoxyethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

To a mixture of 1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde(For an example preparation see Intermediate 2, 1 g, 6.62 mmol) andsodium hydrosulfite (3.46 g, 19.85 mmol) was added a solution of(4-((2-methoxyethyl)amino)-3-nitrophenyl)methanol (For an examplepreparation see Intermediate 199, 1.497 g, 6.62 mmol) in ethanol (30mL), followed by water (15 mL). The reaction mixture was heated at 80°C. overnight. The reaction mixture was partitioned between saturatedaqueous sodium hydrogen carbonate solution (150 mL) and 3:1chloroform:isopropanol (3×150 mL). The organic layers were combined,dried using a hydrophobic frit and evaporated under reduced pressure.The sample was loaded in dichloromethane and purified by SPE (silica,100 g) using a gradient of 0-12% (2 M ammonia in methanol) in DCM. Theappropriate fractions were combined and evaporated under reducedpressure to give the title compound (2.0 g, 6.11 mmol) as an off-whitefoam. LCMS (System B): t_(RET)=0.62 min, MH⁺=328.

Intermediate 203:2-(1,5-Dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxyethyl)-1H-benzo[d]imidazole-5-carbaldehyde

To a solution of5-(5-(hydroxymethyl)-1-(2-methoxyethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(For an example preparation see Intermediate 202, 2.0 g, 6.11 mmol) inDCM (50 mL) was added 45% iodoxybenzoic acid (stabilised by benzoic acidand isophthalic acid) (4.18 g, 6.72 mmol), portionwise. The reactionmixture was stirred at room temperature for 2 nights. The reactionmixture was partitioned between 10% isopropanol in dichloromethane (200mL) and saturated aqueous sodium bicarbonate solution (3×200 mL). Theorganic layer was dried using a hydrophobic frit and evaporated underreduced pressure to give the title compound (975 mg, 3.00 mmol) as apale yellow solid. LCMS (System B): t_(RET)=0.71 min, MH⁺=326.

Intermediate 204:(4-((2-(Dimethylamino)ethyl)amino)-3-nitrophenyl)methanol

A mixture of (4-fluoro-3-nitrophenyl)methanol (2.5 g, 14.61 mmol),N1,N1-dimethylethane-1,2-diamine (3.19 mL, 29.2 mmol) and DIPEA (7.65mL, 43.8 mmol) in THF (20 mL) in two equal portions was under microwaveconditions (initial high absorption setting) at 120° C. for 30 min. Thereaction mixture was partitioned between dichloromethane (3×150 mL) andsaturated aqueous sodium bicarbonate solution (150 mL). The organiclayers were combined, dried using a hydrophobic frit and evaporatedunder reduced pressure. The sample was loaded in dichloromethane andpurified by SPE (silica, 100 g). The appropriate fractions were combinedand evaporated under reduced pressure to give the title compound (3.32g, 13.88 mmol) as a dark orange gum. LCMS (System B): t_(RET)=0.73 min,MH⁺=240.

Intermediate 205:5-(1-(2-(Dimethylamino)ethyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

To a mixture of 5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (Foran example preparation see Intermediate 1, 3.3 g, 24.06 mmol) and sodiumhydrosulfite (7.20 g, 41.4 mmol) was added a solution of(4-((2-(dimethylamino)ethyl)amino)-3-nitrophenyl)methanol (For anexample preparation see Intermediate 204, 3.3 g, 13.79 mmol) in ethanol(26 mL), followed by water (13 mL). The reaction mixture was heated, intwo equal portions, under microwave conditions (initial high absorptionsetting) at 100° C. for 5 hours. The reaction mixture was partitionedbetween saturated aqueous sodium hydrogen carbonate solution (100 mL)and dichloromethane (2×100 mL), followed by further extraction of theaqueous layer with 3:1 chloroform:isopropanol (3×100 mL). The organiclayers were combined and evaporated under reduced pressure. The samplewas loaded in methanol/dichloromethane (and the column dried in a vacuumoven) and purified by SPE (silica, 100 g) using a gradient of 0-50% (2 Mammonia in methanol) in DCM. The appropriate fractions were combined andevaporated under reduced pressure. The sample was dried in a vacuum ovento give the title compound (1.75 g, 5.36 mmol). LCMS (System B):t_(RET)=0.58 min, MH⁺=327.

Intermediate 206:1-(2-(Dimethylamino)ethyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde

To a solution of5-(1-(2-(dimethylamino)ethyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(For an example preparation see Intermediate 205, 1.31 g, 4.01 mmol) inDCM (50 mL) was added 45% iodoxybenzoic acid (stabilised by benzoic acidand isophthalic acid) (2.75 g, 4.41 mmol), portionwise. The reactionmixture was stirred at room temperature for 5 nights. Further 45%iodoxybenzoic acid (stabilised by benzoic acid and isophthalic acid)(1.38 g, 2.21 mmol) was added, followed by DMSO (10 mL). The reactionmixture was stirred at room temperature for a further 7 nights. Thereaction mixture was partitioned between 10% isopropanol indichloromethane (150 mL) and saturated aqueous sodium bicarbonatesolution (3×150 mL). The organic layers were combined, dried using ahydrophobic frit and blown down under a stream of nitrogen to give thetitle compound (758 mg, 2.337 mmol) as an off-white solid. LCMS (SystemB): t_(RET)=0.66 min, MH⁺=325.

Intermediate 207:5-(5-(Hydroxymethyl)-1-methyl-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

(4-(Methylamino)-3-nitrophenyl)methanol (For an example preparation seeIntermediate 1, 5.32 g, 29.2 mmol),5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (For an examplepreparation see Intermediate 71, 4.00 g, 29.2 mmol) and sodiumdithionite (15.25 g, 88 mmol) were dissolved in ethanol (75 mL), water(37.5 mL) and refluxed over 14 h. Thereby, the mixture was partitionedthrough the addition of 200 mL of saturated sodium bicarbonate and 200mL of ethyl acetate. The organic phase was isolated and the aqueousextracted twice with 200 mL of ethyl acetate. The organics werecombined, dried over magnesium sulfate, solvent removed. The residue wastriturated with diethyl ether and filtered to give the title compound(1.519 g, 5.64 mmol) as a pale yellow solid. LCMS (System A):t_(RET)=0.39 min, MH⁺=270.

Intermediate 208:1-Methyl-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde

To a solution of5-(5-(hydroxymethyl)-1-methyl-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(For an example preparation see Intermediate 207, 1.88 g, 6.98 mmol) inDCM (50 mL) was added 45% iodoxybenzoic acid (stabilised by benzoic acidand isophthalic acid) (4.78 g, 7.68 mmol), portionwise. The reactionmixture was stirred at room temperature for 4 nights. DMSO (10 mL) wasadded and the reaction mixture stirred at room temperature for a further1 night. The reaction mixture was partitioned between dichloromethane(100 mL) and saturated aqueous sodium bicarbonate solution (3×100 mL).The organic layer was dried using a hydrophobic frit and evaporatedunder reduced pressure to give the title compound (655 mg, 2.451 mmol)as a dark brown solid. LCMS (System B): t_(RET)=0.61 min, MH⁺=268.

Intermediate 209:(3-Nitro-4-(((tetrahydro-2H-pyran-3-yl)methyl)amino)phenyl)methanol

A solution of (tetrahydro-2H-pyran-3-yl)methanamine (4.11 mL, 32.1mmol), (4-fluoro-3-nitrophenyl)methanol (2200 mg, 12.86 mmol) andN-ethyl-N-isopropylpropan-2-amine (11.23 mL, 64.3 mmol) in THF (30 mL)was degassed and heated under nitrogen at 60° C. overnight. The solventwas removed under reduced pressure and the residue was partitionedbetween ethyl acetate (150 mL) and a saturated solution of sodiumbicarbonate (150 mL). The organic fraction was isolated and the aqueouslayer was re-extracted twice with ethyl acetate (2×150 mL). The organicfractions were combined, passed through a hydrophobic frit andconcentrated under reduced pressure. The residue was dissolved in aminimum amount of dichloromethane purified by SPE (silice, 2×100 g),eluted with a gradient of 0-70% EtOAc in cyclohexane. The productcontaining fractions were combined and concentrated under reducedpressure to yield the title compound (2731 mg) as an orange solid. LCMS(System B): t_(RET)=0.82 min, MH⁺=267.

Intermediate 210:5-(5-(Hydroxymethyl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

A solution of(3-nitro-4-(((tetrahydro-2H-pyran-3-yl)methyl)amino)phenyl)methanol (Foran example preparation see Intermediate 1, 2600 mg, 9.76 mmol),5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (For an examplepreparation see Intermediate 209, 1339 mg, 9.76 mmol) and sodiumdithionite (5100 mg, 29.3 mmol) in ethanol (30 mL) and water (15 mL) wasdegassed. The reaction mixture was heated under nitrogen at 80° C.overnight. The reaction mixture was allowed to cool down to roomtemperature and then partitioned between 3:1 chloroform:isopropanol (150mL) and a saturated solution of sodium bicarbonate (150 mL). The organicfraction was isolated and the aqueous layer was re-extracted twice with3:1 chloroform:isopropanol (2×150 mL). The organic fractions werecombined, passed through a hydrophobic frit and concentrated underreduced pressure. The residue was dissolved in a minimum amount of 10%methanol in ethyl acetate. The solution was loaded onto two SPE columns(100 g, silica) and the solvents in which the product was loaded ontothe columns allowed to evaporate. The product was then eluted with agradient of 0-30% EtOH in EtOAc. The product containing fractions werecombined and concentrated under reduced pressure to yield the titlecompound (1500 mg) as a white solid. LCMS (System B): t_(RET)=0.63 min,MH⁺=354.

Intermediate 211:2-(5-Methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

A suspension of 45% 2-iodoxybenzoic acid (stabilised by benzoic andisonapthalic acids) (6603 mg, 10.61 mmol) and5-(5-(hydroxymethyl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(For an example preparation see Intermediate 210, 1500 mg, 4.24 mmol)was stirred at room temperature for 3 days under nitrogen. The reactionmixture was partitioned between 3:1 chloroform:isopropanol (75 mL) and asaturated solution of sodium bicarbonate (75 mL). The organic layer wasisolated and the aqueous fraction was re-extracted twice with 3:1chloroform:isopropanol (2×75 mL). The organic fractions were combined,passed through a hydrophobic frit and concentrated under reducedpressure. The material was partitioned between 3:1chloroform:isopropanol (75 mL) and saturated solution of sodiumbicarbonate (75 mL). The organic layer was isolated and the aqueousfraction was re-extracted twice with 3:1 chloroform:isopropanol (2×75mL). The organic fractions were combined, passed through a hydrophobicfrit, concentrated under reduced pressure, and dried under vacuum for 7days to yield the title compound (1487 mg) as a light pink solid. LCMS(System B): t_(RET)=0.70 min, MH⁺=352.

Intermediate 212:2-(1,5-Dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

A solution of2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(For an example preparation see Intermediate 211, 700 mg, 1.992 mmol) inDMF (10 mL) was treated with potassium carbonate (551 mg, 3.98 mmol) andthe reaction mixture was stirred for 2 hours. Iodomethane (0.149 mL,2.390 mmol) was added and the mixture was stirred overnight. The solventwas removed under reduced pressure and the residue was partitionedbetween 3:1 chloroform:isopropanol (75 mL) and a saturated solution ofsodium bicarbonate (75 mL). The organic layer was isolated and theaqueous fraction was re-extracted twice with 3:1 chloroform:isopropanol(2×75 mL). The organic fractions were combined, passed through ahydrophobic frit and concentrated under reduced pressure. The resultinggum was dissolved in a minimum amount of 10% methanol in EtOAc andloaded onto an SPE column (50 g, silica). The product was eluted with agradient of 0-30% EtOH in EtOAc. Product containing fractions werecombined and concentrated under reduced pressure to yield the titlecompound (451 mg) as a brown solid. LCMS (System A): t_(RET)=0.73 min,MH⁺=366.

Intermediate 213:(S)-(4-((1-Methoxypropan-2-yl)amino)-3-nitrophenyl)methanol

To a solution of (4-fluoro-3-nitrophenyl)methanol (2 g, 11.69 mmol) inTHF (10 mL) was added (S)-1-methoxypropan-2-amine (1.851 mL, 17.53 mmol)and DIPEA (6.12 mL, 35.1 mmol) and the reaction mixture was heated undermicrowave conditions (initial high absorption setting) at 120° C. for 30min. The reaction mixture was partitioned between dichloromethane (3×150mL) and saturated aqueous sodium bicarbonate solution (150 mL). Theorganic layers were combined, dried using a hydrophobic frit andevaporated under reduced pressure. The sample was loaded indichloromethane and purified by SPE (silica, 100 g) using a gradient of0-25% EtOAc in cyclohexane. The appropriate fractions were combined andevaporated under reduced pressure to give the title compound (1.4 g,5.83 mmol) as a orange gum. LCMS (System B): t_(RET)=0.82 min, MH⁺=241.

Intermediate 214:(S)-5-(5-(Hydroxymethyl)-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

To a mixture of 1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde(For an example preparation see Intermediate 2, 0.881 g, 5.83 mmol) andsodium hydrosulfite (3.04 g, 17.48 mmol) was added a solution of(S)-(4-((1-methoxypropan-2-yl)amino)-3-nitrophenyl)methanol (For anexample preparation see Intermediate 213, 1.4 g, 5.83 mmol) in ethanol(30 mL), followed by Water (15 mL). The reaction mixture was heated at80° C. overnight. The reaction mixture was partitioned between saturatedaqueous sodium hydrogen carbonate solution (150 mL) and ethyl acetate(3×150 mL), followed by 3:1 chloroform:isopropanol (3×150 mL). Theorganic layers were combined, dried using a hydrophobic frit andevaporated under reduced pressure. The sample was loaded indichloromethane and purified by SPE (silica, 100 g) using a gradient of0-25% EtOAc in cyclohexane. The appropriate fractions were combined andevaporated under reduced pressure to give the title compound (580 mg,1.699 mmol) as a off-white foam. LCMS (System B): t_(RET)=0.67 min,MH⁺=342

Intermediate 215:(S)-2-(1,5-Dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazole-5-carbaldehyde

To a solution of(S)-5-(5-(hydroxymethyl)-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(For an example preparation see Intermediate 214, 577 mg, 1.690 mmol) inDCM (25 mL) was added 45% iodoxybenzoic acid (stabilised by benzoic acidand isophthalic acid) (1157 mg, 1.859 mmol), portionwise. The reactionmixture was stirred at room temperature for 3 nights. The reactionmixture was partitioned between 10% isopropanol in dichloromethane (200mL) and saturated aqueous sodium bicarbonate solution (3×200 mL). Theorganic layer was dried using a hydrophobic frit and evaporated underreduced pressure to give the title compound (630 mg, 1.856 mmol) as anoff-white foam. LCMS (System B): t_(RET)=0.76 min, MH⁺=340.

Intermediate 216:(R)-(4-((1-Methoxypropan-2-yl)amino)-3-nitrophenyl)methanol

To a solution of (4-fluoro-3-nitrophenyl)methanol (5 g, 29.2 mmol) inTHF (40 mL) was added (R)-1-methoxypropan-2-amine hydrochloride (5.50 g,43.8 mmol) and DIPEA (22.96 mL, 131 mmol) and the reaction mixture washeated at 60° C. overnight. The reaction mixture was partitioned betweendichloromethane (3×250 mL) and saturated aqueous sodium bicarbonatesolution (150 mL). The organic layers were combined, dried using ahydrophobic frit and evaporated under reduced pressure. The sample wasloaded in dichloromethane and purified by SPE (silica, 2×100 g), using agradient of 0-25% EtOAc in cyclohexane. The appropriate fractions werecombined and evaporated under reduced pressure to give the titlecompound (1.7 g, 7.08 mmol) as an orange gum. LCMS (System B):t_(RET)=0.81 min, MH⁺=241.

Intermediate 217:(R)-5-(5-(Hydroxymethyl)-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

To a mixture of 1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde(For an example preparation see Intermediate 2, 0.629 g, 4.16 mmol) andsodium hydrosulfite (2.174 g, 12.49 mmol) was added a solution of(R)-(4-((1-methoxypropan-2-yl)amino)-3-nitrophenyl)methanol (For anexample preparation see Intermediate 216, 1 g, 4.16 mmol) in ethanol (30mL), followed by water (15 mL). The reaction mixture was heated at 90°C. for 6 hours. The reaction mixture was partitioned between saturatedaqueous sodium hydrogen carbonate solution (150 mL) and ethyl acetate(3×150 mL), followed by 3:1 chloroform:isopropanol (3×150 mL). Theorganic layers were combined, dried using a hydrophobic frit andevaporated under reduced pressure. The sample was loaded indichloromethane and purified by SPE (silica, 100 g) using a gradient of0-100% EtOAc in cyclohexane, followed by a gradient of 0-100% (25% EtOHin EtOAc) in cyclohexane. The appropriate fractions were combined andevaporated under reduced pressure to give the title compound (393 mg,1.151 mmol) as an off-white foam. LCMS (System B): t_(RET)=0.67 min,MH⁺=342.

Intermediate 218:(R)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazole-5-carbaldehyde

To a solution of(R)-5-(5-(hydroxymethyl)-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(For an example preparation see Intermediate 217, 390 mg, 1.142 mmol) inDCM (25 mL) was added 45% iodoxybenzoic acid (stabilised by benzoic acidand isophthalic acid) (782 mg, 1.257 mmol), portionwise. The reactionmixture was stirred at room temperature for 3 nights. The reactionmixture was partitioned between 3:1 chloroform:IPA (200 mL) andsaturated aqueous sodium bicarbonate solution (3×200 mL). The organiclayer was dried using a hydrophobic frit and evaporated under reducedpressure. The sample was dissolved in 3:1 chloroform:IPA (200 mL) andwashed with saturated aqueous sodium bicarbonate solution (3×200 mL).The organic layer was dried using a hydrophobic frit and evaporatedunder reduced pressure to give the title compound (554 mg, 1.632 mmol,143% yield), in approximately 70% purity. LCMS (System B): t_(RET)=0.76min, MH⁺=340.

Intermediate 219:(3-Nitro-4-(((tetrahydrofuran-3-yl)methyl)amino)phenyl)methanol

(Tetrahydrofuran-3-yl)methanamine (1.711 mL, 16.36 mmol) was dissolvedin THF (5 mL), and DIPEA (5.00 mL, 28.6 mmol) and(4-fluoro-3-nitrophenyl)methanol (1400 mg, 8.18 mmol) were added. Thereaction mixture was heated for 2.5 h under microwave conditions at 125°C. The reaction mixture was partitioned between a saturated solution ofsodium bicarbonate (100 mL) and ethyl acetate (100 mL). The organiclayer was isolated and the aqueous fraction was extracted twice withethyl acetate (2×100 mL). The organic fractions were combined, passedthrough a hydrophobic frit and concentrated under reduced pressure. Theresidue was dissolved in a minimum amount of DCM and purified by SPE(silica, 100 g), eluted with a gradient of 0-65% EtOAc in cyclohexane.The purest fractions were combined and concentrated under reducedpressure to give the title compound (1719 mg) as an orange gum. LCMS(System B): t_(RET)=0.78 min, MH⁺=253.

Intermediate 220:5-(5-(Hydroxymethyl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(3-Nitro-4-(((tetrahydrofuran-3-yl)methyl)amino)phenyl)methanol (For anexample preparation see Intermediate 219, 1719 mg, 6.81 mmol) wasdissolved in ethanol (16.00 mL) and water (8 mL) and1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (For an examplepreparation see Intermediate 2, 1145 mg, 6.81 mmol) was added. Thereaction mixture was heated under nitrogen until it reached the reactiontemperature (80° C.) and sodium dithionite (3559 mg, 20.44 mmol) wasadded. The reaction mixture was stirred under nitrogen for 4 hours. Thereaction mixture was concentrated under reduced pressure and the residuewas partitioned between 10% IPA/DCM (100 mL) and a saturated solution ofsodium bicarbonate (100 mL). The organic layer was isolated, the aqueousfraction was extracted with 10% IPA/DCM (2×100 mL). The organicfractions were combined, passed through a hydrophobic frit andconcentrated under reduced pressure. The residue was dissolved in aminimum amount of DCM and purified by SPE (100 g, silica), eluted with agradient of 0-25% EtOH in EtOAc. The appropriate fractions were combinedand concentrated under reduced pressure to give the title compound (1189mg) as a white solid. LCMS (System B): t_(RET)=0.65 min, MH⁺=354.

Intermediate 221:2-(1,5-Dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

5-(5-(Hydroxymethyl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(For an example preparation see Intermediate 220, 1189 mg, 3.36 mmol)was dissolved in DCM (20 mL). IBX (45 wt %, stabilised by benzoic andnaphthalic acids) (5234 mg, 8.41 mmol) was added and the reactionmixture was stirred for 3 days. The reaction mixture was partitionedbetween 10% IPA/DCM (50 mL) and a saturated solution of sodiumbicarbonate (50 mL). The organic layer was isolated and the aqueousfraction was extracted twice with 10% IPA/DCM (2×50 mL). The organicfractions were combined and washed with a saturated solution of sodiumbicarbonate (75 mL). The organic layer was isolated, passed through ahydrophobic frit and concentrated under reduced pressure. The residuewas placed in a vacuum oven for 30 minutes to give the title compound(1271 mg) as a white solid. LCMS (System B): t_(RET)=0.73 min, MH⁺=352.

Intermediate 222:(4-((1,3-Dimethoxypropan-2-yl)amino)-3-nitrophenyl)methanol

To a solution of (4-fluoro-3-nitrophenyl)methanol (16 g, 93 mmol) in THF(80 mL) was added 1,3-dimethoxypropan-2-amine (10 g, 84 mmol) and DIPEA(15 mL, 86 mmol). The mixture was split into seven portions and heatedunder microwave conditions at 100° C. for 7 hours. The reaction mixturewas partitioned between ethyl acetate (3×500 mL) and saturated aqueoussodium bicarbonate solution (750 mL). The organic layers were combined,washed with saturated brine (500 mL), and evaporated under reducedpressure. The sample was loaded in dichloromethane and purified SPE(silica, 100 g) using a gradient of 0-80% EtOAc in cyclohexane. Theappropriate fractions were combined and evaporated under reducedpressure to give a the title compound (12.0 g, 44.4 mmol). LCMS (SystemB): t_(RET)=0.85 min, MH⁺=271. Less pure fractions were combined andevaporated. The sample was loaded in dichloromethane and purified by SPE(silica, 100 g) using a gradient of 0-50% EtOAc in cyclohexane. Theappropriate fractions were combined and evaporated under reducedpressure to give the title compound (3.7 g, 13.69 mmol). LCMS (SystemJ): t_(RET)=0.85 min, MH⁺=271.

Intermediate 223:5-(1-(1,3-Dimethoxypropan-2-yl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

To a solution of(4-((1,3-dimethoxypropan-2-yl)amino)-3-nitrophenyl)methanol (For anexample preparation see Intermediate 222, 12 g, 44.4 mmol) in ethanol(160 mL) was added sodium hydrosulfite (27.5 g, 133 mmol) and1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (For an examplepreparation see Intermediate 2, 8.39 g, 55.5 mmol), followed by water.The reaction mixture was heated at 70° C. for two hours The reactionmixture was concentrated to approximately half the volume under reducedpressure, and the resulting liquid partitioned between 3:1chloroform:isopropanol (3×300 mL) and saturated aqueous sodiumbicarbonate solution (300 mL). The organic layers were combined, driedusing a hydrophobic frit and evaporated under reduced pressure. Crudematerial from a similar reaction (an additional 40% scale) was added atthis stage. The sample was loaded in DCM and purified by SPE (silica,3×340 g) using a gradient of 0-100% (25% EtOH in EtOAc) in cyclohexane.The appropriate fractions were combined and evaporated under reducedpressure to give the title compound (7.5 g, 20.19 mmol) as an off-whitefoam. LCMS (System J): t_(RET)=0.70 min, MH⁺=372.

Intermediate 224:1-(1,3-Dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-benzo[d]imidazole-5-carbaldehyde

To a solution of5-(1-(1,3-dimethoxypropan-2-yl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(For an example preparation see Intermediate 223, 7.5 g, 20.19 mmol) inDCM (400 mL) was added 45% iodoxybenzoic acid (stabilised by benzoicacid and isophthalic acid) (13.82 g, 22.21 mmol) and the reactionmixture stirred at room temperature for 3 nights. Saturated aqueoussodium bicarbonate solution (400 mL) was added slowly, and the mixturestirred until bubbling had stopped. The organic layer was removed, andthe aqueous reextracted with 3:1 chloroform:isopropanol (2×400 mL). Theorganic layers were combined, dried using a hydrophobic frit andevaporated under reduced pressure to give the title compound (7.3 g,19.76 mmol). LCMS (System J): t_(RET)=0.81 min, MH⁺=370.

Intermediate 225:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

5-(5-(hydroxymethyl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 226) (600 mg, 1.633 mmol) andDess-Martin periodinane (693 mg, 1.633 mmol) were dissolved in DCM (5mL) and stirred under nitrogen for 2 hrs. Dess-Martin periodinane (291mg, 0.544 mmol) was then added and the mixture stirred for a further 30min. Saturated sodium bicarbonate solution (25 mL) was then added andthe mixture stirred for 25 minutes then left standing overnight. To themixture was added dichloromethane (25 mL) and the phases separated. Theaqueous phase was then extracted with dichloromethane (25 mL×2) and theorganics combined. The organic phase was then washed with brine (30mL×2) and the organics combined and dried over a hydrophobic frit.Solvents were then removed in vacuo yielding the title compound (550 mg,1.505 mmol, 92% yield) as an off-white solid. LCMS (System B):t_(RET)=0.87 min; MH⁺ 366.

Intermediate 226:5-(5-(hydroxymethyl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(3-Nitro-4-(((tetrahydro-2H-pyran-2-yl)methyl)amino)phenyl)methanol (fora preparation see Intermediate 227, 2.9 g, 10.89 mmol),1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, 2.469 g, 16.34 mmol) and sodium dithionite (5.69 g,32.7 mmol) were added to flask along with ethanol (35 mL). This mixturewas heated to 90° C. and stirred for 15 minutes. Water (35 mL) was thenadded and the mixture stirred under nitrogen for 20 hours. The mixturewas allowed to cool to room temperature. The mixture was thenpartitioned between 3:1 dichloromethane:isopropyl alcohol (100 mL) andwater (100 mL) and the phases separated. Aqueous phase was extractedwith 3:1 dichloromethane:isopropyl alcohol (100 mL×2) and the organicscombined. Organic phase was then dried using a hydrophobic frit andsolvents removed in vacuo yielding the title compound (1.99 g, 5.42mmol, 49.7% yield) as an off-white solid. LCMS (System B): t_(RET)=0.77min; MH⁺ 368.

Intermediate 227:(3-nitro-4-(((tetrahydro-2H-pyran-2-yl)methyl)amino)phenyl)methanol

(4-fluoro-3-nitrophenyl)methanol (1.985 g, 11.60 mmol) and(tetrahydro-2H-pyran-2-yl)methanamine (4.44 mL, 34.7 mmol) weredissolved in THF (10 mL) and N,N-Diisopropylethylamine (6.08 mL, 34.8mmol) added. Mixture was heated in a microwave reactor at 120° C. for 1hr. The orange mixture was then partitioned between ethyl acetate (100mL) and saturated sodium bicarbonate (100 mL). The phases were separatedand the aqueous layer extracted twice with ethyl acetate (100 mL). Theorganics were then combined, dried using a hydrophobic frit and solventsremoved in vacuo. This yielded the title compound as a bright orangesolid. Excessively high yield must be due to a misweighing of a startingmaterial. LCMS (System B): t_(RET)=0.92 min; MH⁺ 267.

Intermediate 228:2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-ethyl-1H-benzo[d]imidazol-5-yl)oxy)acetaldehyde

Dess-Martin periodinane (143 mg, 0.34 mmol) was added portionwise to astirred solution of5-(1-ethyl-5-(2-hydroxyethoxy)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 229, 100 mg, 0.31 mmol) indichloromethane (10 mL). The reaction mixture was stirred at roomtemperature under nitrogen for 4 hours. A further portion of Dess-Martinperiodinane (143 mg, 0.34 mmol) was added portionwise and the reactionmixture was stirred at room temperature overnight. 10% sodiumthiosulphate solution (20 mL) was added. The organic phase was separatedand the aqueous phase was extracted with dichloromethane (2×10 mL). Thecombined extracts were washed with saturated NaHCO₃ solution, dried andevaporated. The residue title compound (99 mg, 0.305 mmol, 100% yield)was used in the next steps without further purification. 100% yieldassumed. LCMS (System B): t_(RET)=0.67 min; M+18⁺ 344.

Intermediate 229:5-(1-ethyl-5-(2-hydroxyethoxy)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

A mixture of5-(1-ethyl-5-hydroxy-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 230, 600 mg, 2.12 mmol),2-bromoethanol (291 mg, 165 μL, 2.33 mmol) and potassium carbonate (878mg, 6.35 mmol) in DMF (3 mL) was heated at 80° C. for 3 days. Thereaction mixture was allowed to cool to room temperature and partitionedbetween ethyl acetate (25 mL) and water (15 mL). The organic phase wasseparated, dried and evaporated. The residue was chromatographed [0-10%ethanol/ethyl acetate] to give the title compound (105 mg, 0.321 mmol,15.15% yield), as an off-white solid. LCMS (System B): t_(RET)=0.66 min;MH⁺ 328.

Intermediate 230:5-(1-ethyl-5-hydroxy-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

Sodium dithionite (2.72 g, 15.64 mmol) was added to a mixture of4-(ethylamino)-3-nitrophenol (for a preparation see Intermediate 231,950 mg, 5.21 mmol) and1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, 985 mg, 6.52 mmol) in ethanol (20 mL) and water (10mL). The reaction mixture was stirred at 80° C. overnight. The reactionmixture was cooled to room temperature and the ethanol was evaporated.Ethyl acetate (100 mL) and saturated NaHCO₃ solution (50 mL) were addedto the residue. A solid was formed. The solid was filtered off and driedto give the title compound (1.2 g, 4.24 mmol, 81% yield), as acolourless solid. LCMS (System A): t_(RET)=0.46 min; MH⁺ 284.

Intermediate 231: 4-(ethylamino)-3-nitrophenol and Intermediate 232:3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenol

A mixture of 4-amino-3-nitrophenol (5.06 g, 32.9 mmol),tetrahydro-2H-pyran-4-carbaldehyde (3.0 g, 26.3 mmol), and acetic acid(1.97 g, 1.88 mL, 32.9 mmol) in dichloromethane (100 mL) was stirred atroom temperature for 1 hour. Sodium triacetoxyborohydride (16.71 g, 79mmol) was added over 5 minutes. After complete addition the reactionmixture was stirred at room temperature for 24 hours. Saturated NaHCO₃solution (100 mL) was added carefully (gas evolved) and the mixturestirred at room temperature for 30 minutes. The organic phase wasseparated. The aqueous phase was extracted with dichloromethane (3×50mL). The combined organics were dried and evaporated. The residue waschromatographed [0-15% ethanol/ethyl acetate] to give;4-(ethylamino)-3-nitrophenol (950 mg, 5.21 mmol, 19.84% yield), as anorange solid (unexpected product from reduction of acetanilide formed byreaction of triacetoxyborohydride with starting material). LCMS (SystemA): t_(RET)=0.87 min; MH⁺ 183. Plus N31507-32-A3,3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenol (450 mg, 1.784mmol, 6.79% yield), as an orange solid. LCMS (System A): t_(RET)=0.85min; MH⁺ 253.

Intermediate 233:(R)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-hydroxypropan-2-yl)-1H-benzo[d]imidazole-5-carbaldehyde

To(R)-5-(5-(hydroxymethyl)-1-(1-hydroxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 234, 500 mg, 1.527 mmol) andDess-Martin Periodinane (648 mg, 1.527 mmol) was added dichloromethane(DCM) (20 mL). Mixture was stirred under nitrogen for 20 hrs; LCMSshowed remaining starting material so Dess-Martin Periodinane (65 mg,0.157 mmol) was added and the mixture stirred for 3 hrs. To the mixturewas added saturated sodium bicarbonate (100 mL) and it was stirred for30 min. Dichloromethane (80 mL) was then added and the phases separated.The aqueous phase was then extracted with dichloromethane (100 mL×2).The combined organics were then washed with brine (100 mL) then driedusing a hydrophobic frit. Solvents were then removed in vacuo yieldingthe title compound (460 mg, 1.414 mmol, 93%) as a yellow solid. LCMS(System A): t_(RET)=0.65 min; MH⁺ 326.

Intermediate 234:(R)-5-(5-(hydroxymethyl)-1-(1-hydroxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(R)-2-((4-(hydroxymethyl)-2-nitrophenyl)amino)propan-1-ol (for apreparation see Intermediate 235, 1.902 g, 8.41 mmol) and1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, 1.906 g, 12.61 mmol) were dissolved in ethanol (30mL) and water (15 mL) and heated to 90° C. for 15 minutes. Sodiumdithionite (4.39 g, 25.2 mmol) was then added and the mixture stirredfor 20 hours under nitrogen. The mixture was then allowed to cool toroom temperature. The mixture was then partitioned between 2:1dichloromethane:isopropanol (75 mL) and water (70 mL). Phases wereseparated and the aqueous phase extracted with 2:1dichloromethane:isopropanol (75 mL×2). Organics were then dried over ahydrophobic frit then solvents removed in vacuo. This yielded a yellowoil which was dissolved in dichloromethane and loaded onto a SNAP 100 gSilica column. This was eluted running a gradient of 0%-30%ethanol/ethyl acetate. Appropriate fractions were combined and solventsremoved in vacuo to give the title compound (1.139 g, 3.48 mmol, 41.4%yield) as a white solid. LCMS (System B): t_(RET)=0.58 min; MH⁺ 328.

Intermediate 235:(R)-2-((4-(hydroxymethyl)-2-nitrophenyl)amino)propan-1-ol

(4-fluoro-3-nitrophenyl)methanol (1.5 g, 8.77 mmol) and(R)-2-aminopropan-1-ol (2.047 mL, 26.3 mmol) were dissolved intetrahydrofuran (THF) (5 mL) and N,N-diisopropylethylamine (4.59 mL,26.3 mmol) was added. Mixture was then heated in a microwave reactor at120° C. for 1 hr. The orange mixture was then partitioned between ethylacetate (80 mL) and saturated sodium bicarbonate (80 mL) and the phasesseparated. Aqueous phase was then extracted with ethyl acetate (80 mL×2)and the organics combined. The organic phase was then dried using ahydrophobic frit then solvents removed in vacuo to give the titlecompound (1.904 g, 8.41 mmol, 96% yield) as an orange/red solid. LCMS(System B): t_(RET)=0.63 min; MH⁺ 227.

Intermediate 236:(S)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-hydroxypropan-2-yl)-1H-benzo[d]imidazole-5-carbaldehyde

(S)-5-(5-(hydroxymethyl)-1-(1-hydroxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 237, 808 mg, 2.468 mmol) andDess-Martin periodinane (1047 mg, 2.468 mmol) were dissolved in DCM (30mL) and the mixture stirred for 3 hours. Dess-Martin Periodinane (104.7mg, 0.247 mmol) was added. The mixture was then stirred overnight thensodium hydroxide solution (2M, 5 mL) and saturated sodium bicarbonatesolution (20 mL) were then added and the mixture stirred for 3 hrs. Tothe mixture was then added dichloromethane (60 mL) and water (60 mL) andthe phases separated. The aqueous phase was then extracted three timeswith dichloromethane (60 mL) and the organics combined. Combinedorganics were then dried using a hydrophobic frit, then solvents removedin vacuo. This yielded the title compound (592 mg, 1.82 mmol, 74%) as ayellow solid. LCMS (System B): t_(RET)=0.64 min; MH⁺ 326.

Intermediate 237:(S)-5-(5-(hydroxymethyl)-1-(1-hydroxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(S)-2-((4-(hydroxymethyl)-2-nitrophenyl)amino)propan-1-ol (for apreparation see Intermediate 238, 1.8 g, 7.96 mmol),1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, 1.323 g, 8.75 mmol) and sodium dithionite (4.16 g,23.87 mmol) were dissolved in ethanol (30 mL) and water (15 mL) and themixture heated to 90° C. for 20 hours. The mixture was then partitionedbetween dichloromethane:isopropanol (2:1; 100 mL) and water (100 mL) andthe phases separated. Aqueous phase was then extracted twice withdichloromethane:isopropanol (2:1; 100 mL) and the organics combined anddried. The solvents were then removed in vacuo. Residue was thendissolved in dichloromethane and loaded onto a Biotage SNAP 100 g Silicacolumn and eluted using a gradient of 0%-25% ethanol/ethyl acetate.Appropriate fractions were then combined and solvents removed in vacuo.This yielded the title compound (808 mg, 2.468 mmol, 31.0% yield) as awhite solid. LCMS (System A): t_(RET)=0.40 min; MH⁺ 328.

Intermediate 238:(S)-2-((4-(hydroxymethyl)-2-nitrophenyl)amino)propan-1-ol

4-fluoro-3-(nitrophenyl)methanol (2 g, 11.69 mmol) was dissolved intetrahydrofuran (5 mL) and DIPEA (6.12 mL, 35.1 mmol) and(S)-2-aminopropan-1-ol (2.73 mL, 35.1 mmol) added. This mixture washeated to 120° C. in a microwave reactor for 1 hour. The orange mixturewas then partitioned between ethyl acetate (80 mL) and saturated sodiumbicarbonate (80 mL). Phases were then separated and the aqueous phasewas then extracted twice with ethyl acetate (80 mL). Organics were thencombined, dried using a hydrophobic frit then solvents removed in vacuo.To the oil was added cyclohexane and the mixture left to stand for 16hrs. Mixture was then stirred for 2.5 hrs then the solvent was decanted.The mixture was then dissolved in dichloromethane/methanol then solventsremoved in vacuo. This yielded the title compound (1.8 g, 7.96 mmol,68.1% yield) as a red/orange oil. LCMS (System B): t_(RET)=0.63 min; MH⁺328.

Intermediate 239:2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-6-yl)oxy)acetaldehyde

Dess-Martin periodinane (1.12 g, 2.64 mmol) was added portionwise to astirred solution of5-(6-(2-hydroxyethoxy)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 240, 700 mg, 1.76 mmol) indichloromethane (25 mL). After complete addition the reaction mixturewas stirred at room temperature for 24 hours. 10% aq. Sodiumthiosulphate solution (25 mL) was added and the mixture stirred for 15minutes. The organic phase was separated. The aqueous phase wasextracted with dichloromethane (2×20 mL). The combined organics werewashed with saturated NaHCO₃ solution (20 mL). The organic phase wasdried and evaporated to give the title compound (696 mg, 1.761 mmol,100% yield), as a light brown oil. LCMS (System B): t_(RET)=0.57 min;M+18⁺ 414.

Intermediate 240:5-(6-(2-hydroxyethoxy)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

A mixture of5-(6-hydroxy-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 241, 2.0 g, 5.66 mmol),2-bromoethanol (778 mg, 442 μL, 6.22 mmol) and potassium carbonate(2.346 g, 16.98 mmol) in dry DMF (15 mL) was stirred at 110° C. for 24hours. The reaction mixture was cooled to room temperature, thenpartitioned between ethyl acetate (50 mL) and water (50 mL). The organicphase was separated and the aqueous phase was extracted with ethylacetate (3×25 mL). The combined organics were dried and evaporated. Theresidue was chromatographed [5-25% ethanol/ethyl acetate] to give thetitle compound (720 mg, 1.811 mmol, 32.0% yield), as a colourless solid.LCMS (System B): t_(RET)=0.63 min; MH⁺ 398.

Intermediate 241:5-(6-hydroxy-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

Sodium dithionite (7.04 g, 40.4 mmol) was added portionwise to a stirredmixture of 4-nitro-3-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenol(for a preparation see Intermediate 242, 3.4 g. 13.48 mmol) and1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, 2.45 g, 16.21 mmol) in ethanol (50 mL) and water (25mL). After complete addition the reaction mixture was refluxed for 4hours. The reaction mixture was cooled and the ethanol was evaporated.The residue was partitioned between ethyl acetate (50 mL) and water (50mL). The organic phase was separated and the aqueous phase was extractedwith ethyl acetate (3×25 mL). The combined organics were dried andevaporated. The residue was chromatographed [5-20% ethanol/ethylacetate] to give the title compound (3.77 g, 10.67 mmol, 79% yield), asa colourless solid. LCMS (System B): t_(RET)=0.61 min; MH⁺ 354.

Intermediate 242:4-nitro-3-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenol

A mixture of 3-fluoro-4-nitrophenol (3.0 g, 19.1 mmol),(tetrahydro-2H-pyran-4-yl)methanamine (3.30 g, 28.6 mmol), anddiisopropylethylamine (3.70 g, 5.0 mL, 28.6 mmol) in dioxan (50 mL) wasrefluxed for 3 hours. The reaction mixture was cooled to roomtemperature and the solvent was evaporated. The residue waschromatographed [50-75% ethyl acetate/cyclohexane] to give the titlecompound (3.42 g, 13.56 mmol, 71.0% yield), as an orange solid. LCMS(System A): t_(RET)=0.86 min; MH⁺ 253.

Intermediate 243:5-(1-(2-hydroxyethyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, 1.726 g, 11.42 mmol),2-((4-(hydroxymethyl)-2-nitrophenyl)amino)ethanol for a preparation seeIntermediate 244, 2.203 g, 10.38 mmol) and sodium dithionite (5.42 g,31.1 mmol) were dissolved in ethanol (30 mL) and water (15 mL) andstirred at 90° C. for 48 hours. Mixture was then allowed to cool to roomtemperature then 3:1 DCM/isopropanol (100 mL) and water (100 mL) added.Phases were separated then the aqueous phase extracted twice with 3:1DCM/isopropanol (100 mL). Organics were combined then dried using ahydrophobic frit then solvents removed in vacuo. The yellow residueproduced was then loaded onto a Biotage SNAP 100 g silica column theneluted with 0%-25% ethanol/ethyl acetate. Appropriate fractions werecombined and solvents removed in vacuo. This yielded the title compound(975 mg, 3.11 mmol, 30.0% yield) as a white solid. LCMS (System B):t_(RET)=0.57 min; MH⁺ 314.

Intermediate 244: 2-((4-(hydroxymethyl)-2-nitrophenyl)amino)ethanol

(4-fluoro-3-nitrophenyl)methanol (2 g, 11.69 mmol) was dissolved intetrahydrofuran (5 mL) then 2-aminoethanol (2.120 mL, 35.1 mmol) andN,N-Diisopropylethylamine (6.12 mL, 35.1 mmol) were added. This mixturewas heated in a microwave reactor at 120° C. for 1 hour. LCMS analysisshowed presence of starting material and the sample was heated in themicrowave reactor at 120° C. for 30 min. Mixture was then partitionedbetween ethyl acetate (75 mL) and saturated sodium bicarbonate (75 mL)and the phases separated. Aqueous phase was then extracted twice withethyl acetate (75 mL) and the organics combined and retained. Theaqueous phase was then extracted with 1:1 mixture ofisopropanol/dichloromethane (75 mL) and the organic phase combined withthe previous organics. Organics were dried over a hydrophobic frit andsolvents removed in vacuo. This yielded the title compound (2.203 g,10.38 mmol, 89% yield) as an orange/red solid. LCMS (System A):t_(RET)=0.57 min; MH⁺ 213.

Intermediate 245:2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

To5-(5-(hydroxymethyl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(for a preparation see Intermediate 246, 1046 mg, 2.96 mmol) andDess-Martin periodinane (1255 mg, 2.96 mmol) was added Dichloromethane(20 mL) and the mixture stirred for 20 hours. LCMS analysis of the brownmixture showed presence of starting material therefore Dess-MartinPeriodinane (126 mg, 0.296 mmol) was added and the mixture stirred for 3hours. To the mixture was added saturated sodium bicarbonate solution(100 mL) and dichloromethane (80 mL) and the phases separated. Organicswere then extracted twice with dichloromethane (100 mL) and organicscombined. Organics were then washed with brine (100 mL) then dried usinga hydrophobic frit. Solvents were removed in vacuo producing the crudetitle compound (1.2 g, 3.41 mmol, 115% yield) as an off-white solid withsome evidence of remaining Dess-Martin Periodinane residues. Nopurification attempted as product is an intermediate. Compound appearedas a pale brown solid. LCMS (System A): t_(RET)=0.76 min; MH⁺ 352.

Intermediate 246:5-(5-(hydroxymethyl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

(3-nitro-4-(((tetrahydro-2H-pyran-2-yl)methyl)amino)phenyl)methanol (fora preparation see Intermediate 227, 2.7 g, 10.14 mmol) and5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparation seeIntermediate 1, 1.808 g, 13.18 mmol) were dissolved in ethanol (35 mL)and water (35.0 mL) and heated to 90° C. Sodium dithionite (5.30 g, 30.4mmol) was then added and the mixture stirred for 20 hours. Mixture wasallowed to cool to room temperature then partitioned between 3:1dichloromethane:isopropanol (160 mL) and sodium bicarbonate (160 mL) andphases separated. Organic phase was then extracted twice with 3:1dichloromethane:isopropanol (160 mL). The organics were then combined,dried using a hydrophobic frit and solvents removed in vacuo. Thisyielded a yellow residue which solidified after being last to stand inair. An attempt was made to dissolve the sample in 25% ethanol/ethylacetate, however, the solid was quite insoluble. The mixture was thusfiltered under gravity and the residue washed with 25% ethanol/ethylacetate. The solid was dried in air then dried further in vacuum oven at40° C. for 1 hour to give the title compound (1.05 g, 2.97 mmol, 29.3%yield) as an off white solid. LCMS (System A): t_(RET)=0.52 min; MH⁺354.

Intermediate 247:(S)-2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazol-5-yl)oxy)acetaldehyde

Dess-Martin periodinane (320 mg, 0.75 mmol) was added portionwise to astirred solution of(S)-5-(5-(2-hydroxyethoxy)-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 248, 140 mg, 0.38 mmol) indichloromethane (10 mL). The reaction mixture was stirred at roomtemperature overnight. 10% sodium thiosulphate solution (20 mL) wasadded and the mixture stirred for 15 minutes. The organic phase wasseparated. The aqueous phase was extracted with dichloromethane (2×10mL). The combined organics were washed with saturated NaHCO₃ (10 mL),dried and evaporated to give the title compound (139 mg, 0.377 mmol,100% yield), as a colourless gum. Quantitative yield assumed used innext steps without further purification. LCMS (System B): t_(RET)=0.66min; M+18⁺ 388.

Intermediate 248:(S)-5-(5-(2-hydroxyethoxy)-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

A mixture of(S)-5-(5-hydroxy-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 249, 1.25 g, 3.82 mmol),2-bromoethanol (525 mg, 298 ⋅L, 4.2 mmol) and potassium carbonate (1.58g, 11.45 mmol) in DMF (50 mL) was stirred at 130° C. for 48 hours. Thereaction mixture was cooled and the solvent was evaporated. The residuewas suspended in ethyl acetate (50 mL) and filtered through ‘celite’.The solvent was evaporated from the filtrate. The residue was purifiedby silica gel column chromatography [10-25% ethanol/ethyl acetate] togive the title compound (140 mg, 0.377 mmol, 9.87% yield), as a yellowgum. LCMS (System B): t_(RET)=0.75 min; MH⁺ 372.

Intermediate 249:(S)-5-(5-hydroxy-1-(1-methoxypropan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

A solution of (S)-4-((1-methoxypropan-2-yl)amino)-3-nitrophenol (for apreparation see Intermediate 250, 1.3 g, 5.75 mmol) and1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, N-Me pyridone aldehyde, 1.04 g, 6.90 mmol) inethanol (30 mL) and water (15 mL) was treated with sodium dithionite(3.00 g, 17.24 mmol) added portion wise over 5 minutes. After completeaddition the reaction mixture was refluxed for 4 hours. The reactionmixture was allowed to cool to room temperature. The ethanol wasevaporated. The residue was partitioned between ethyl acetate (50 mL)and water (25 mL). The organic phase was separated. The aqueous phasewas extracted with ethyl acetate (2×25 mL). The combined organics weredried and evaporated. The residue was chromatographed [5-20%ethanol/ethyl acetate] to give the title compound (1.27 g, 3.88 mmol,67.5% yield), as an off white solid. LCMS (System A): t_(RET)=0.46 min;MH⁺ 328.

Intermediate 250: (S)-4-((1-methoxypropan-2-yl)amino)-3-nitrophenol

A mixture of 4-fluoro-3-nitrophenol (1.0 g, 6.37 mmol),(S)-1-methoxypropan-2-amine (1.13 g, 1.35 mL, 12.73 mmol) anddiisopropylethylamine (1.65 g, 2.22 mL, 12.73 mmol) in dioxan (10 mL)was refluxed for 2 hours. The solvent was evaporated, the residue wasdissolved in N-methyl-2-pyrrolidone (10 mL). The mixture was heated in amicrowave at 180° C. for 4 hours. The cooled reaction mixture waspartitioned between ethyl acetate (50 mL) and water (25 mL). The organicphase was separated, washed with water (2×25 mL), dried and evaporated.The residue was chromatographed [10-40% ethyl acetate/cyclohexane] togive the title compound (1.31 g, 5.79 mmol, 91% yield), as an orangesolid. LCMS (System A): t_(RET)=0.89 min; MH⁺ 227.

Intermediate 251:2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(Enantiomer 1, a Single Enantiomer of Unknown Configuration)

Dess-Martin periodinane (625 mg, 1.47 mmol) was added portionwise to astirred solution of5-(5-(hydroxymethyl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(Enantiomer 1, for a preparation see Intermediate 252, 250 mg, 0.74mmol) in dichloromethane (10 mL). The reaction mixture was stirred atroom temperature overnight. 10% sodium thiosulphate solution (20 mL) wasadded and the mixture stirred for 15 minutes. The organic phase wasseparated. The aqueous phase was extracted with dichloromethane (2×10mL). The combined organics were washed with saturated NaHCO₃ solution(10 mL), dried and evaporated to give the title compound (249 mg, 0.737mmol, 100% yield), as a colourless gum. Quantitative yield assumed. LCMS(System B): t_(RET)=0.67 min; MH⁺ 338.

Intermediate 252:5-(5-(hydroxymethyl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(Enantiomer 1) and Intermediate 253:5-(5-(hydroxymethyl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(Enantiomer 2) Chiral Separation of Intermediate 254:

Analytical Method: Approx 0.5 mg dissolved in 50% EtOH/Heptane (1 mL) 20ul injected on column (40% EtOH/Heptane, f=1.0 mL/min, wavelength 215nm, 4. Ref 550,100, Column 4.6 mmid×25 cm Chiralpak IA)

Prep Method: Approx 2.25 g dissolved in 12 mL of EtOH+heat. Injection; 2mL of the solution was injected onto the column (40% EtOH/Heptane, f=30mL/min, wavelength 215 nm, 4.

Ref 550,100, Column 30 mm×25 cm Chiralpak IA)

Intermediate 252 (Enantiomer 1) Rt=11.5 min. >99% ee by UV.

Intermediate 253 (Enantiomer 2) Rt=16.5 min. >99% ee by UV.

Intermediate 254:5-(5-(hydroxymethyl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one

Sodium dithionite (7.66 g, 44.0 mmol) was added in three single portionsto a suspension of(3-nitro-4-(((tetrahydrofuran-3-yl)methyl)amino)phenyl)methanol (for apreparation see Intermediate 219, 3.7 g, 14.67 mmol) and5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparation seeIntermediate 1, 2.51 g, 15.56 mmol) in ethanol (60 mL) and water (30mL). The reaction mixture was stirred at 80° C. overnight. The reactionmixture was cooled to room temperature and the ethanol was evaporated.The mixture was partitioned between DCM:iPrOH 3:1 (150 mL) andNaHCO₃(100 mL). The separated aqueous phase was extracted with DCM:iPrOH3:1 (3×150 mL). The combined organic phases were passed through ahydrophobic frit and evaporated to obtain a pale yellow oil. The samplewas loaded in 20% MeOH/DCM and purified by chromatography (Biotage SP4)on SNAP 100 g silica column using a 10-20% MeOH/DCM gradient. Theappropriate fractions were combined and dried down to give the titlecompound (2.37 g, 6.98 mmol, 48%) as a pale yellow oil. LCMS (System C):t_(RET)=0.42 min; MH⁺ 340.

Intermediate 255:2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(Enantiomer 2)

Dess-Martin periodinane (625 mg, 1.47 mmol) was added portionwise to astirred solution of5-(5-(hydroxymethyl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(Enantiomer 2, for a preparation see Intermediate 253, 250 mg, 0.74mmol) in dichloromethane (10 mL). The reaction mixture was stirred atroom temperature overnight. 10% sodium thiosulphate solution (20 mL) wasadded and the mixture stirred for 15 minutes. The organic phase wasseparated. The aqueous phase was extracted with dichloromethane (2×10mL). The combined organics were washed with saturated NaHCO₃ solution(10 mL), dried and evaporated to give the title compound (249 mg, 0.737mmol, 100% yield), as a colourless gum. Quantitative yield assumed. Usedwithout further purification in next steps. LCMS (System B):t_(RET)=0.67 min; MH+338.

Intermediate 256:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1H-benzo[d]imidazole-5-carbaldehyde

5-(5-(hydroxymethyl)-1-(2-methoxypropyl)-H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 257, 2.153 g, 6.31 mmol) wasdissolved in dichloromethane (15 mL) then Dess-Martin periodinane (2.67g, 6.31 mmol) was added. Mixture was stirred under nitrogen for 1 hour.To the mixture was added sodium bicarbonate (200 mL) and dichloromethane(200 mL). This mixture was stirred for 1.5 hours then phase separationattempted. The organics from the separation were retained. 2M HCl (100mL) was added. The organic phase was separated and dried using ahydrophobic frit, and solvents removed in vacuo. This yielded a brownresidue, the crude title compound. LC-MS purity was ˜45%. LCMS (SystemA): t_(RET)=0.73 min; MH⁺ 340.

Intermediate 257:5-(5-(hydroxymethyl)-1-(2-methoxypropyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

(4-((2-methoxypropyl)amino)-3-nitrophenyl)methanol (for a preparationsee Intermediate 258, 2.8152 g, 11.72 mmol) and1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 2, 1.948 g, 12.89 mmol) were dissolved in ethanol (50mL) and water (25 mL) and heated to 90° C. under nitrogen for 1 hour.Sodium dithionite (8.16 g, 46.9 mmol) was then added and the mixturestirred under nitrogen for 1.5 hours at 90° C. The mixture was thenpartitioned between saturated sodium bicarbonate (100 mL) and 3:1dichloromethane:isopropanol (100 mL) and the phases separated. Aqueousphase was then extracted twice with 3:1 dichloromethane:isopropanol (100mL) the organics were combined then dried over a hydrophobic frit.Solvents were then removed in vacuo leaving a yellow residue. Residuewas dissolved in dichloromethane, split into two portions and loadedonto two Biotage SNAP 100 g Silica columns. Each column was elutedrunning a gradient of 0%-25% ethanol/ethyl acetate. Appropriatefractions were then combined and solvents removed in vacuo. This yieldedthe title compound (2.1253 g, 6.23 mmol, 53.1% yield) as a brown solid.LCMS (System A): t_(RET)=0.48 min; MH⁺ 342.

Intermediate 258: (4-((2-methoxypropyl)amino)-3-nitrophenyl)methanol

(4-fluoro-3-nitrophenyl)methanol (2.72 g, 15.89 mmol),2-methoxypropan-1-amine, hydrochloride (2.99 g, 23.84 mmol), anddiisopropylethylamine (13.88 mL, 79 mmol) were added to tetrahydrofuran(10 mL) and split into two equal portions. Each portion was placed in a20 mL microwave vial and heated to 120° C. for 1 hour in a microwavereactor. Starting material was still present therefore sample was heatedin the microwave reactor for 30 minutes at 120° C. The mixtures werecombined and partitioned between ethyl acetate (150 mL) and saturatedsodium bicarbonate (150 mL). Aqueous phase was then extracted twice withethyl acetate (150 mL), organics combined and dried using a hydrophobicfrit. Solvents were then removed in vacuo yielding a red oil. The oilwas dissolved in dichloromethane and loaded onto two Biotage SNAP 100 gSilica columns. Each column was eluted using a gradient of 10%-50% ethylacetate/cyclohexane. Appropriate fractions were combined yielding thetitle compound (2.8152 g, 11.72 mmol, 73.7% yield) as an orange solid.LCMS (System A): t_(RET)=0.82 min; MH⁺ 241.

Intermediate 259:5-(5-hydroxy-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenol (for apreparation see Intermediate 232, 3.68 g, 14.59 mmol),1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for a preparationsee Intermediate 1, 2.205 g, 14.59 mmol) and sodium hydrosulfite (7.62g, 43.8 mmol) were stirred in Ethanol (64.8 mL) under nitrogen. Water(32.4 mL) was added and the reaction mixture was stirred at 90° C. undernitrogen overnight. The reaction mixture was cooled down to roomtemperature and concentrated under reduced pressure to ⅓ of its volume.The residue was taken in DCM and washed with water. The organics weredried over a phase separator and concentrated under reduced pressure.The red solid was purified by silica gel column chromatography elutedwith 0-25% EtOH in ethyl acetate. The fractions were concentrated underreduced pressure to give the title compound (3.48 g). LCMS (System B):t_(RET)=0.67 min, MH⁺=354.

Intermediate 260:5-(5-(2-hydroxyethoxy)-1-((tetrahydro-2-pyran-4-yl)methyl)-1-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

5-(5-hydroxy-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 259, 0.75 g, 2.122 mmol),2-bromoethanol (0.318 g, 2.55 mmol) and K₂CO₃ (0.880 g, 6.37 mmol) indry DMF (7.07 mL) were stirred at 120° C. under nitrogen. After 18 hrs,2-bromoethanol (0.318 g, 2.55 mmol) was added and the reaction mixturewas stirred at 120° C. for 7 hrs. The mixture was allowed to cool downto room temperature. Water and ethyl acetate were added and the layerswere separated. The organics were washed with water, dried through ahydrophobic frit and concentrated under reduced pressure. The residuewas purified by flash column chromatography eluted with 5-25% ethanol inethyl acetate to give the title compound as a white solid (0.18 g). LCMS(System B): t_(RET)=0.70 min, MH⁺=398.

Intermediate 261:2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)oxy)acetaldehyde

5-(5-(2-hydroxyethoxy)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 260, 0.18 g, 0.453 mmol) wasdissolved in DCM (8 mL) under nitrogen atmosphere. Dess-martinperiodinane (0.288 g, 0.679 mmol) was added. The reaction mixture wasstirred at room temperature overnight. A saturated solution of sodiumthiosulfate (20 mL) was added and the mixture was stirred at roomtemperature for 15 min. The mixture was diluted with DCM and the layersseparated. The aqueous was extracted with DCM. The organics werecombined and washed with a saturated sodium bicarbonate solution, driedthrough a phase separator and concentrated to give the title compound asa yellow solid. Further experiments were carried out on the crudematerial. LCMS (System B): t_(RET)=0.65 min, MH⁺=396.

Intermediate 262: (2S,3R)-isopropyl2-(2-(4-fluoro-3-nitrophenyl)acetamido)-3-hydroxybutanoate

2-(4-fluoro-3-nitrophenyl)acetic acid (2 g, 10.04 mmol),(2S,3R)-isopropyl 2-amino-3-hydroxybutanoate, 4-Methylbenzenesulphonicacid salt (for a preparation see Intermediate 31, 4.02 g, 12.05 mmol),pyridine (2.437 mL, 30.1 mmol) and n-propylphosphonic acid anhydride,cyclic trimer (8.88 mL, 15.07 mmol) in Ethyl acetate (45 mL) werestirred at room temperature overnight. Pyridine (2.437 mL, 30.1 mmol)was added and the reaction mixture further stirred at room temperaturefor 18 hrs. The reaction mixture was diluted with water. The layers wereseparated. The aqueous were extracted with ethyl acetate. The organicswere washed with brine and then water, dried over a phase separator andconcentrated. The residue was purified by flash column chromatographyeluted with 0-10% ethanol in ethyl acetate. The fractions wereconcentrated to give the title compound (31% yield). LCMS (System B):t_(RET)=0.90 min, MH⁺=343.

The following Intermediate was prepared in a similar manner tointermediate 262:

Intermediate 263: (2S,3R)-tert-butyl2-(2-(4-fluoro-3-nitrophenyl)acetamido)-3-hydroxybutanoate

(prepared from commercially available starting materials) System B, 0.96min, MH⁺=355.

Intermediate 264: (2S,3R)-isopropyl3-hydroxy-2-(2-(3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)acetamido)butanoate

(2S,3R)-isopropyl2-(2-(4-fluoro-3-nitrophenyl)acetamido)-3-hydroxybutanoate (for apreparation see Intermediate 262, 1.51 g, 4.41 mmol), DIPEA (2.311 mL,13.23 mmol) and (tetrahydro-2H-pyran-4-yl)methanamine (0.762 g, 6.62mmol) in THF (40 mL) were stirred under nitrogen at 60° C. overnight.DIPEA (2.311 mL, 13.23 mmol) and (tetrahydro-2H-pyran-4-yl)methanamine(0.2 mL) were added and the reaction mixture was heated at 60° C. fortwo hours. The reaction mixture was concentrated under reduced pressure.The residue was taken in DCM and washed with water. The organics weredried over a phase separator and concentrated to give an orange oil. Theoil was purified by flash column chromatography eluted with 25-100%ethyl acetate in cyclohexane. The fractions were concentrated to givethe title compound (58% yield).

LCMS (System B): t_(RET)=0.95 min, MH⁺=438.

The following Intermediate was prepared in a similar way to Intermediate264:

Intermediate 265: (2S,3R)-tert-butyl3-hydroxy-2-(2-(3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)acetamido)butanoate

(prepared from: Intermediate 263) System B, 1.00 min, MH⁺=452.

Intermediate 266:1,3-dimethyl-5-(5-(oxiran-2-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)pyridin-2(1-H)-one

2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(for a preparation see Intermediate 116, 0.100 g, 0.274 mmol),trimethylsulfonium iodide (0.057 g, 0.279 mmol) and potassium hydroxide(0.092 g, 1.642 mmol) were stirred in Acetonitrile (2.72 mL) and Water(0.014 mL) under nitrogen at 65° C. for 4 hours. The reaction was cooleddown to room temperature, diluted with ethyl acetate and water. Theorganics were washed with sodium bicarbonate. The organics were thendried over a phase separator and concentrated under reduced pressure togive the title compound as a white solid (49% yield). LCMS (System B):t_(RET)=0.79 min, MH⁺=389.

Intermediate 267: (R)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-((((2S,3R)-3-hydroxy-1-isopropoxy-1-oxobutan-2-yl)amino)methyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate

Sodium triacetoxyborohydride (68 mg, 0.33 mmol) was added to a stirredsolution of (R)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-formyl-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate(for a preparation see Intermediate 126, 100 mg, 0.22 mmol) and(2S,3R)-isopropyl 2-amino-3-hydroxybutanoate 4-methylbenzenesulfonate(For a preparation see Intermediate 31, 86 mg, 0.26 mmol). The resultingsuspension was stirred overnight. Further (2S,3R)-isopropyl2-amino-3-hydroxybutanoate 4-methylbenzenesulfonate (86 mg, 0.26 mmol)and Sodium triacetoxyborohydride (68 mg, 0.33 mmol) were added and thesuspension stirred for 2 h. The reaction mixture was solubilised withMeOH and loaded on to a 5 g SCX cartridge. The cartridge was eluted withMeOH, followed by 2M methanolic ammonia. The basic fractions wereevaporated in vacuo to a colourless gum and purified by MDAP (Method B).The product containing fractions were evaporated in vacuo to give thetitle compound (29 mg) as a white foam. LCMS (System C): t_(RET)=0.66min, MH⁺=610.

Intermediate 268: (R)-tert-butyl3-((6-((((2S,3R)-1-(tert-butoxy)-3-hydroxy-1-oxobutan-2-yl)amino)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate

Sodium triacetoxyborohydride (46 mg, 0.22 mmol) was added to a stirredsolution of (R)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-formyl-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate(For a preparation see Intermediate 126, 50 mg, 0.11 mmol) and(2S,3R)-tert-butyl 2-amino-3-hydroxybutanoate.HCl (28 mg, 0.13 mmol).The reaction mixture was stirred overnight. (2S,3R)-tert-butyl2-amino-3-hydroxybutanoate hydrochloride (28 mg, 0.13 mmol) was added,the resulting suspension stirred for 2 h and sodiumtriacetoxyborohydride (46 mg, 0.22 mmol) added. The resulting suspensionwas stirred for 1 h. The suspension was solubilised with MeOH and loadedon to a 5 g SCX cartridge. The cartridge was eluted with MeOH, followedby 2M methanolic ammonia. The basic fractions were evaporated in vacuoto a colourless gum and purified by MDAP (Method B). The productcontaining fractions were evaporated in vacuo to give the title compound(20 mg) as a white foam.

LCMS (System C): t_(RET)=0.69 min, MH⁺=624.

Intermediate 269: (S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-((((S)-1-isopropoxy-3-methyl-1-oxobutan-2-yl)amino)methyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate

(S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-formyl-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate(For a preparation see Intermediate 128, 70 mg, 0.151 mmol) wasdissolved in DCM (3 mL) and (S)-isopropyl 2-amino-3-methylbutanoate4-methylbenzenesulfonate (For a preparation see Intermediate 65,N31501-74-A2, 100 mg, 0.301 mmol) was added. The suspension was stirredunder nitrogen flow for 1 hr. Sodium triacetoxyborohydride (63.9 mg,0.301 mmol) was added and the reaction mixture was stirred at rt for 23hr. (S)-isopropyl 2-amino-3-methylbutanoate 4-methylbenzenesulfonate(49.9 mg, 0.151 mmol) was added and stirred for 30 min prior to additionof sodium triacetoxyborohydride (31.9 mg, 0.151 mmol). The mixture wasstirred at rt for 3.5 hr. The reaction mixture was diluted with DCM (20mL) and NaHCO₃(20 mL) was added. The separated aqueous phase wasextracted with DCM (3×20 mL). The combined organic phases were passedthrough a hydrophobic frit and evaporated to obtain the crude product(124 mg) as a yellow solid. The sample was dissolved in MeOH 2 mL andpurified by MDAP (Method B). The solvent was dried down to give thetitle compound (65 mg) as a yellow-orange solid. LCMS (System B):t_(RET)=1.30 min, MH⁺=608.

Intermediate 270: (S)-tert-butyl3-((5-(((1-(cyclopentyloxy)-4-methyl-1-oxopentan-2-yl)amino)methyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-1-yl)methyl)azetidine-1-carboxylate

A microwave vial was charged with (S)-tert-butyl3-(((4-(((1-(cyclopentyloxy)-4-methyl-1-oxopentan-2-yl)amino)methyl)-2-nitrophenyl)amino)methyl)azetidine-1-carboxylate(for a preparation see Intermediate 271, 355 mg, 0.684 mmol), THF (12mL), 5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for apreparation see Intermediate 1, 94 mg, 0.684 mmol) and a solution ofsodium hydrosulfite (417 mg, 2.396 mmol) in Water (3 mL). The vial wascapped and the mixture heated in a microwave using initial normal to100° C. for 5 hours. The organic solvent was removed in vacuo andreplaced with ethanol (12 mL). The vial was capped and the mixtureheated in a microwave to 100° C. for a total of 5 hours. The mixture wasdiluted with EtOAc and water, the layers mixed and separated before theorganics were passed through a hydrophobic frit and concentrated invacuo to give a yellow oil. The oil was loaded in dichloromethane andpurified by Biotage SP4 SNAP 25 g silica using a gradient of 0-100%ethyl acetate-cyclohexane over 15 CV followed by 0-20% MeOH-DCM over 15CV. The appropriate fractions were combined and evaporated in vacuo togive the title compound (60 mg, 0.099 mmol, 14.47% yield) as a yellowoil. LCMS (System A): t_(RET)=0.88 min; MH⁺ 606.

Intermediate 271: (S)-tert-butyl3-(((4-(((1-(cyclopentyloxy)-4-methyl-1-oxopentan-2-yl)amino)methyl)-2-nitrophenyl)amino)methyl)azetidine-1-carboxylate

A round bottom flask was charged with (S)-cyclopentyl2-((4-fluoro-3-nitrobenzyl)amino)-4-methylpentanoate (for a preparationsee Intermediate 137, 230 mg, 0.653 mmol), acetonitrile (10 mL),potassium carbonate (180 mg, 1.305 mmol) and tert-butyl3-(aminomethyl)azetidine-1-carboxylate (146 mg, 0.783 mmol). An aircondenser was fitted and the mixture warmed to 80° C. under a blanket ofnitrogen for 1 week. The mixture was diluted with water and ethylacetate, the layers mixed and separated before the organics were washedwith brine. The organics were passed through a hydrophobic frit andconcentrated in vacuo to give an orange oil. The oil was loaded indichloromethane and purified by Biotage SP4 SNAP 25 g silica (Si) usinga gradient of 0-40% ethyl acetate-cyclohexane over 15 CV. Theappropriate fractions were combined and evaporated in vacuo to give thetitle compound (355 mg, 0.684 mmol, 105% yield) as an orange oil. LCMS(System A): t_(RET)=1.07 min; MH⁺ 519.

Intermediate 272: (S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-5-((((S)-1-isopropoxy-3-methyl-1-oxobutan-2-yl)amino)methyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate

(S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-5-formyl-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate(for an example preparation see intermediate 130, 500 mg, 1.076 mmol)was added to a solution of (S)-isopropyl 2-amino-3-methylbutanoate,phenylmethane sulphonic acid salt (for a preparation see Intermediate65, 713 mg, 2.153 mmol) in Dichloromethane (DCM) (10 mL). The resultingsolution was stirred overnight and sodium triacetoxyborohydride (684 mg,3.23 mmol) added. The resulting suspension was stirred for 1 h. MeOH (10mL) was added and the resulting solution loaded on to a 10 g SCXcartridge. The cartridge was eluted with MeOH (50 mL), followed by 2 Mmethanolic ammonia (50 mL). The basic fractions were evaporated in vacuoto a brown gum. The gum was dissolved in DCM and purified by silica gelchromatography eluting with DCM:2M methanolic ammonia (0-5%). Theproduct containing fractions were evaporated in vacuo to give the titlecompound. The total yield of the reaction was 28%. LCMS (System C):t_(RET)=0.75 min, MH⁺=608.

Intermediate 273:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-methyl-1H-benzo[d]imidazole-5-carbaldehyde

5-(5-(hydroxymethyl)-1-methyl-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 90, 931 mg, 3.29 mmol) was dissolvedin dichloromethane (DCM) (52 mL). To the stirred solution was added2-iodoxybenzoic acid (2249 mg, 3.61 mmol). The mixture was stirred undernitrogen at room temperature for 2.5 hrs then1,1,1-Triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one1,1,1-Triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one (1394 mg, 3.29mmol) was added portion wise to the stirred solution. The mixture wasstirred at room temperature overnight. The solution was then partitionedbetween saturated sodium bicarbonate (200 mL) and dichloromethane (100mL), the phases separated and the aqueous phase extracted twice withdichloromethane (100 mL×2). The organics were then combined and driedusing a hydrophobic frit and the solvent removed under vacuum. To theresidue was added a few mLs of dichloromethane followed by a few mLs of2M NaOH and the solution was stirred for 15 min. The mixture was thenpartitioned between dichloromethane (50 mL) and saturated sodiumbicarbonate (50 mL) and the phases separated taking care not to allowany of the precipitated solid to enter the organic phase. The aqueousphase was then washed twice with dichloromethane (2×50 mL) and theorganics combined. The solution was then dried with a hydrophobic fritand the solvent removed under vacuum producing2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-methyl-1H-benzo[d]imidazole-5-carbaldehyde(438 mg, 1.557 mmol, 47.4% yield) as a pale yellow solid. Theprecipitated solid was isolated by filtration and dried under suction toyield an additional batch of2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-methyl-1H-benzo[d]imidazole-5-carbaldehyde(200 mg, 0.710 mmol, 21.6% yield) as a light brown solid. LCMS (SystemB): t_(RET)=0.66 min, MH⁺=282.

Intermediate274:1-ethyl-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde

5-(5-(hydroxymethyl)-1-isopropyl-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 91, 722 mg, 2.319 mmol) wasdissolved in dichloromethane (DCM) (5 mL) and then treated with1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one (983 mg, 2.319mmol). The solution was stirred for 1 hr under nitrogen then 2M NaOH (15mL) added and the solution stirred until the mixture became clear (15min). The mixture was then partitioned between dichloromethane (150 mL)and saturated sodium bicarbonate (150 mL) and the phases separated. Theaqueous layer was then extracted twice with saturated sodium bicarbonate(150 mL×2) and the organics combined. The organic layer was then driedusing a hydrophobic frit and solvent removed in vacuo. The residue wasthen purified using automated chromatography using the following method:100% ethyl acetate for 2 column volumes, 0%-25% ethanol in ethyl acetateover 15 column volumes then 25% ethanol in ethyl acetate over 2 columnvolumes. The appropriate fractions were then combined and solventremoved in vacuo to yield2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-isopropyl-1H-benzo[d]imidazole-5-carbaldehyde(718.7 mg, 2.323 mmol, 100% yield) as a white solid. LCMS (System A):t_(RET)=0.70 min, MH⁺=310.

Intermediate 275:2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

A solution of oxalyl chloride (0.124 ml, 1.411 mmol) in dichloromethane(2 mL) was cooled to −78° C. in acetone/dry-ice bath. To this, asolution of DMSO (0.213 ml, 3.01 mmol) in dichloromethane (3 mL) wasadded and the mixture was stirred at −78° C. for 10 minutes. Asuspension of5-(5-(hydroxymethyl)-1-((4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-2-yl)-3-methylpyridin-2(1H)-one(400 mg, 1.086 mmol, for a preparation see intermediate 150) indichloromethane (5 mL) was added and the reaction mixture was stirred at−78° C. for 2 hours. Triethylamine (1.059 ml, 7.60 mmol) was then addedand the reaction mixture was warmed up to room temperature. It wasstirred under nitrogen at room temperature overnight. The reactionmixture was partitioned between saturated sodium hydrogen carbonatesolution (40 mL) and DCM (40 mL) and the layers were separated. Theaqueous layer was extracted with DCM (3×40 mL) and then with 25%methanol in DCM (2×40 mL). The organic layers were combined, dried usinga hydrophobic frit and evaporated under reduced pressure to give acolourless gum. The sample was loaded in dichloromethane and purified byBiotage SP4 SNAP 25 g silica using a gradient of 0-20%dichloromethane-2M ammonia in methanol over 20 column volumes followedby holding at 20% dichloromethane-2M ammonia in methanol for 5 columnvolumes. The pure fractions were combined and evaporated under reducedpressure to give a colourless gum. The less pure fractions containingproduct were combined separately and the solvent was removed underreduced pressure. This impure crude sample was loaded in dichloromethaneand repurified by Biotage SP4 SNAP 25 g silica using a gradient of 4-16%dichloromethane-2M ammonia in methanol over 20 column volumes followedby holding at 16% dichloromethane-2M ammonia in methanol for 5 columnvolumes. The appropriate fractions were combined and evaporated underreduced pressure. This was combined with the pure sample from theearlier run to give the crude product2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(137.8 mg, 0.376 mmol, 34.6% yield) as a pale yellow solid. The samplewas loaded in dichloromethane/methanol and repurified by Biotage SP4SNAP 25 g silica using a gradient of 10-25% methanol-chloroform over 20column volumes followed by holding at 25% methanol-chloroform for 5column volumes. The appropriate fractions were combined and evaporatedunder reduced pressure to give the title compound (53.6 mg, 0.146 mmol,13.47% yield) as a white solid. LCMS (System B): t_(RET)=0.65 min,MH⁺=367.

Intermediate 276: (S)-Cyclopentyl 2-amino-3-hydroxypropanoate,4-methylbenzenesulphonic acid salt

To a suspension of (S)-2-amino-3-hydroxypropanoic acid (15 g, 143 mmol)in cyclohexane (150 mL), cyclopentanol (98 g, 1142 mmol) and4-methylbenzenesulfonic acid (32.0 g, 186 mmol) were added at roomtemperature were added. The reaction mixture was stirred at 100° C. for24 hr. The reaction mixture was evaporated in vacuo to give the crudeproducts as a brown oil. The brown oil was allowed to cool and theresulting crystals filtered, washed with EtOAc (50 mL) to give(S)-cyclopentyl 2-amino-3-hydroxypropanoate, 4-methylbenzenesulphonicacid salt (38 g, 110 mmol, 77% yield) as a white solid. ¹H NMR δ (400MHz, DMSO-d₆) ppm: 8.28 (3H, br. s.), 7.47 (2H, d, J=8.1 Hz), 7.11 (2H,d, J=7.8 Hz), 5.55 (1H, br. s.), 5.19 (1H, t, J=5.7 Hz), 3.81 (1H, dd,J=11.4, 3.8 Hz), 3.73 (1H, dd, J=11.4, 2.8 Hz), 2.29 (3H, s), 1.93-1.78(2H, m), 1.67 (4H, d, J=5.6 Hz), 1.63-1.50 (2H, m).

Intermediate 277: (S)-Cyclopentyl 2-aminopropanoate,4-methylbenzenesulphonic acid salt

A round bottom flask was charged with (S)-2-aminopropanoic acid (20 g,224 mmol), cyclohexane (101 mL), tosic acid monohydrate (55.5 g, 292mmol) and cyclopentanol (166 ml, 1836 mmol). A Dean-Stark condenser wasfitted and the mixture was stirred at 110° C. for 48 h. The reactionmixture was cooled to room temperature and then the solvent wasevaporated in vacuo to give the crude product as a brown liquid. Thecrude product was dissolved in a minimum volume of hot EtOAc (20 mL).The solution was allowed to cool and the resulting crystals filtered,washed with a small amount of EtOAc (10 mL×3) and dried in an oil vacuumto give (S)-cyclopentyl 2-aminopropanoate, 4-methylbenzenesulphonic acidsalt (58 g, 174 mmol, 78% yield) as a white solid. ¹H NMR δ (400 MHz,DMSO-d₆) ppm: 8.23 (3H, br. s.), 7.47 (2H, d, J=8.1 Hz), 7.12 (2H, d,J=7.8 Hz), 5.18 (1H, t, J=5.9 Hz), 4.05 (1H, q, J=6.7 Hz), 2.29 (3H, s),1.92-1.79 (2H, m), 1.72-1.53 (6H, m), 1.36 (3H, d, J=7.3 Hz).

Intermediate 278:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-hydroxyethyl)-1H-benzo[d]imidazole-5-carbaldehyde

To5-(1-(2-hydroxyethyl)-5-(hydroxymethyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 243, 975 mg, 3.11 mmol) andDess-Martin periodinane (660 mg, 1.556 mmol) was added dichloromethane(40 mL). The mixture was stirred for 1.5 hours then Dess-Martinperiodinane (660 mg, 1.556 mmol) was added. The mixture was stirred foran additional 18 hours. Dess-Martin periodinane (66 mg, 0.156 mmol) wasadded and stirred for 2 hours. Dess-Martin periodinane (66 mg, 0.156mmol) was added and stirred for 2 hours. Dess-Martin periodinane (75 mg,0.38 mmol) was then added and the mixture stirred for 2 hours. To themixture was added 2M aq sodium hydroxide solution (20 mL) and themixture stirred for 10 minutes until the mixture became clear. Water (20mL) and dichloromethane (60 mL) were then added and the phasesseparated. Aqueous phase was then extracted with dichloromethane (100mL×2) and the organic phases combined and dried using a hydrophobicfrit. Solvents were then removed in vacuo. This yielded an off-whitesolid2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-hydroxyethyl)-1H-benzo[d]imidazole-5-carbaldehyde(675 mg, 2.168 mmol, 69.7% yield). LCMS (System A): t_(RET)=0.53 min;MH⁺ 312.

Intermediate 279: (S)-(R)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate

A mixture of (S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanoic acid(1.0 g, 4.60 mmol), diisopropylethylamine (1.19 g, 1.608 mL, 9.21 mmol),1-hydroxybenzotriazole hydrate (HOBt) (846 mg, 5.52 mmol), EDC (1.06 g,5.53 mmol), and (R)-1-methoxypropan-2-ol (1.037 g, 1.127 mL, 11.5 mmol)in DMF (5 mL) was stirred at room temperature overnight. The reactionmixture was partitioned between ethyl acetate (15 mL) and sat'dNaHCO₃(15 mL). The organic phase was washed with 1M hydrochloric acid(15 mL), water (15 mL) and brine (15 mL). The organic phase was driedand evaporated under reduced pressure to give the title compound as acolourless oil (1.2 g, 90% yield). ¹H NMR (CDCl₃): δ 0.89 (m, 3H), 0.96(m, 3H), 1.24 (d, J=6.6 Hz, 3H), 1.45 (s, 9H), 2.16 (m, 1H), 3.36 (s,3H), 3.44 (m, 2H), 4.23 (m, 1H), 5.05 (m, 1H), 5.13 (m, 1H).

Intermediate 280: (S)-(R)-1-methoxypropan-2-yl2-amino-3-methylbutanoate, Hydrochloride

(S)-(R)-1-methoxypropan-2-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate (for a preparation seeIntermediate 279, 1.15 g, 3.97 mmol) was dissolved in ethyl acetate (2mL). The solution was treated with 4M hydrogen chloride in dioxan (2mL). The reaction mixture was stirred at room temperature overnight. Thesolvent was evaporated to give the title compound as a colourless gum(425 mg, 47.4% yield). ¹H NMR (CDCl₃): δ 1.15 (m, 6H), 1.29 (d, J=6.4Hz, 3H), 2.47 (m, 1H), 3.35 (s, 3H), 3.47 (m, 2H), 3.95 (m, 1H), 5.18(m, 1H), 8.79 (m, 3H).

Intermediate 281: (S)-cyclopentyl 2-aminopentanoate hydrochloride

(S)-2-aminopentanoic acid (2.5 g, 21.34 mmol) was added to cyclopentanol(30 mL) and the suspension was brought to −5° C. using a dry ice/acetonebath. After stirring at this temperature for ten minutes, thionylchloride (3.58 mL, 49.1 mmol) was added dropwise. The suspension wasleft stirring and allowed to warm up to room temperature. It was thenstirred at this temperature for 18 hours. The reaction mixture waswarmed to 60° C. and stirred at this temperature for 24 hours. Thereaction mixture was concentrated under reduced pressure. The productwas recrystallised from ethyl acetate, filtered, washed and dried togive the title product (2.62 g, 86% yield) as a white solid. ¹H NMR(d₆-DMSO): δ 0.89 (t, J=7.3 Hz, 3H), 1.31 (m, 1H), 1.42 (m, 1H), 1.67(m, 8H), 1.86 (m, 2H), 3.91 (m, 1H), 5.19 (m, 1H), 8.49 (m, 3H).

Intermediate 282: (S)-(S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate

A mixture of (S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanoic acid(2.5 g, 11.51 mmol), diisopropylethylamine (2.97 g, 4.02 mL, 23.01mmol), 1-hydroxybenzotriazole hydrate (2.12 g, 13.84 mmol), EDC (2.65 g,13.81 mmol), and (S)-tetrahydrofuran-3-ol (5.07 g, 3.91 mL, 57.5 mmol)in DMF (20 mL) was stirred at room temperature overnight. The reactionmixture was partitioned between ethyl acetate (50 mL) and saturatedsodium bicarbonate (50 mL). The organic phase was washed with 1 Mhydrochloric acid (50 mL), water (50 mL) and brine (50 mL). The organicphase was dried and evaporated to give the title compound (2.73 g, 9.50mmol, 83% yield), as a colourless oil. ¹H NMR (CDCl₃): δ 0.90 (d, J=7.1Hz, 3H) 0.97 (d, J=6.9 Hz, 3H) 1.45 (s, 9H) 2.01-2.09 (m, 1H) 2.10-2.24(m, 2H) 3.80 (d, J=10.8 Hz, 1H) 3.83-3.97 (m, 3H) 5.04 (d, J=8.8 Hz, 1H)5.27-5.43 (m, 1H).

Intermediate 283: (S)-(S)-tetrahydrofuran-3-yl 2-amino-3-methylbutanoatehydrochloride

A solution of (S)-(S)-tetrahydrofuran-3-yl2-((tert-butoxycarbonyl)amino)-3-methylbutanoate (for an examplepreparation see Intermediate 282, 2.73 g, 9.50 mmol) in ethyl acetate (5mL) was treated with 4 M hydrogen chloride in dioxan (5 mL). Thereaction mixture was stirred at room temperature for 24 hours. Thesolvent was evaporated. Attempted trituration with diethyl ether did notgive solid. The solvent was evaporated to give the title compound (1.98g, 8.85 mmol, 93% yield) as a colourless oil. Sample solidified onstanding at room temperature for several days. ¹H NMR (d₆-DMSO): δ 0.95(d, J=6.9 Hz, 2H) 1.00 (d, J=6.9 Hz, 3H) 1.94-2.02 (m, 1H) 2.13-2.25 (m,2H) 3.71-3.84 (m, 5H) 5.33-5.37 (m, 1H) 8.70 (br.s., 3H).

Intermediate 284:(S)-3-(1-(tert-butoxycarbonyl)-1H-imidazol-4-yl)-2-((tert-butoxycarbonyl)amino)propanoicacid

Solid (S)-2-((tert-butoxycarbonyl)amino)-3-(1H-imidazol-4-yl)propanoicacid (48.27 g, 189 mmol) was dissolved in a solution of sodiumbicarbonate (15.89 g, 189 mmol) in water (200 mL) and 1,4-dioxane (500mL), di-tert-butyl dicarbonate (43.9 mL, 189 mmol) was added at 0° C.The reaction mixture was stirred at 25° C. overnight. The reactionmixture was evaporated and the residue partitioned between water (500mL) and ether (500 mL). Then to the aqueous was added saturated KHSO₄ toadjust pH=4, extracted with ethyl acetate (3×500 mL) and saturated brine(500 mL), dried over sodium sulfate and evaporated in vacuo to give thetitle compound (40 g, 113 mmol, 60% yield) as a white solid. ¹H NMR(CDCl₃): δ 1.38, 1.60 (2×s, total 18H) 3.16, 3.25 (2×m, total 2H) 4.48(m, 1H) 5.54 (m, 1H) 7.21 (s, 1H) 8.17 (s, 1H).

Intermediate 285: (S)-tert-butyl4-(2-((tert-butoxycarbonyl)amino)-3-(cyclopentyloxy)-3-oxopropyl)-1H-imidazole-1-carboxylate

To a solution of(S)-3-(1-(tert-butoxycarbonyl)-1H-imidazol-4-yl)-2-((tert-butoxycarbonyl)amino)propanoicacid (For an example preparation see Intermediate 284, 40 g, 1126 mmol)and cyclopentanol (97 g, 1126 mmol) in DCM (200 mL), DCC (232 g, 1126mmol) and DMAP (13.75 g, 113 mmol) was added. The reaction mixture wasstirred at 25° C. overnight. The reaction was filtered through aseparatory funnel, rinsed with Dichloromethane (DCM) (200 mL), and thefiltrate collected. The filtrate was washed with water (200 mL),saturated brine (200 mL), dried over sodium sulphate and evaporated invacuo to give the crude product. The crude product was added to a silicagel column and was eluted with Hex/EtOAc=4:1. Collected fractions: thetitle compound (36 g, 851 mmol, 75.6% yield) as a brown oil. ¹H NMR(CDCl₃): δ 1.45 (s, 9H) 1.59-1.71 (m, 11H), 1.8 (m, 6H) 3.05 (d, 2H)4.52 (m, 1H) 5.21 (m, 1H) 5.70 (d, 1H) 7.16 (s, 1H) 8.03 (s, 1H).

Intermediate 286: (S)-cyclopentyl 2-amino-3-(1H-imidazol-4-yl)propanoatedihydrochloride

To solid (S)-tert-butyl4-(2-((tert-butoxycarbonyl)amino)-3-(cyclopentyloxy)-3-oxopropyl)-1H-imidazole-1-carboxylate(For an example preparation see Intermediate 285, 36 g, 52.7 mmol), 4 Mhydrochloric acid in ethyl acetate (400 ml, 1600 mmol) was added. Thereaction mixture was stirred at 25° C. for 24 hr. Another hydrochloricin dioxane (400 mL) addition was made and the reaction mixture wasstirred at 25° C. for 24 hr. The reaction mixture was filtered, theresulting solid was filtered through a separatory funnel, rinsed withethyl acetate (25 mL), and collected as a white solid, the titlecompound (14.3 g) which was assumed to be two HCl salt. The filtrate wasevaporated in vacuo to give the crude product. The crude product wastriturated with ethyl acetate (50 mL). The resulting solid was filteredthrough a separatory funnel, rinsed with ethyl acetate (25 mL), andcollected the title compound (7.7 g) as an orange solid, which wasassumed to be the bis hydrochloride salt. ¹H NMR (MeOD): δ 1.61-1.78 (m,6H) 1.95-1.95 (m, 2H) 3.37 (m, 2H) 4.43 (m, 1H) 5.30-5.33 (m, 1H) 7.56(s, 1H) 9.00 (s, 1H).

Intermediate 287: (2R,3S)-isopropyl 2-amino-3-hydroxybutanoatehydrochloride

Acetyl chloride (17.91 mL, 252 mmol) was added dropwise to propan-2-ol(120 mL, 1557 mmol) while cooling in an ice bath, then the mixture wasstirred for 30 min at room temperature. After that(2R,3S)-2-amino-3-hydroxybutanoic acid (10 g, 84 mmol) was added to thereaction mixture and then the reaction mixture was heated at 80° C. for16h. The reaction mixture was concentrated under reduced pressure andthe resultant residue was azeotroped with toluene (2×50 mL), thentriturated with ether (50 mL) and dried to get the title compound(2R,3S)-isopropyl 2-amino-3-hydroxybutanoate hydrochloride (8 g, 48.2%yield) as a white semi solid. LCMS (System G): t_(RET)=1.37 min; MH⁺ 162

Intermediate 288: (2S,3S)-isopropyl 2-amino-3-hydroxybutanoatehydrochloride

Acetyl chloride (4.48 mL, 63.0 mmol) was added dropwise to iPrOH (30.0mL, 389 mmol) while cooling in an ice bath, the mixture was then stirredfor 30 minutes at room temperature before the addition of(2S,3S)-2-amino-3-hydroxybutanoic acid (2.5 g, 20.99 mmol). The mixturewas then heated to 80° C. giving a dense white suspension, whichclarified on further heating to a clear colourless solution. This washeated overnight. The reaction mixture was cooled and evaporated invacuo to give a white solid, the residue was azeotroped with toluene (20mL), then triturated with ether (20 mL), giving the crude title compound(2S,3S)-isopropyl 2-amino-3-hydroxybutanoate hydrochloride (2.69 g, 65%yield) as a white solid. The crude product was used directly in the nextstep without further purification.

¹H NMR (400 MHz, METHANOL-d₄) δ 5.27-5.46 (m, 1H), 4.38-4.59 (m, 2H),4.19-4.27 (m, 1H), 1.29-1.60 (m, 13H (9H from desired compound, plusoverlapping isopropanol impurity)) Overall purity is 66% by NMR.

Intermediate 289: (2R,3R)-isopropyl 2-amino-3-hydroxybutanoatehydrochloride

Acetyl chloride (448 μL, 6.30 mmol) was added dropwise to iPrOH (2999μL, 38.9 mmol) while cooling in an ice bath, the mixture was thenstirred for 30 minutes at room temperature before the addition of(2R,3R)-2-amino-3-hydroxybutanoic acid (250 mg, 2.099 mmol). The mixturewas then heated to 80° C. giving a dense white suspension, whichclarified on further heating to a clear colourless solution. This washeated over the weekend. The reaction mixture was cooled and evaporatedin vacuo to give a white solid, the residue was azeotroped with toluene(20 mL), then triturated with ether (20 mL), giving the title compound(2R,3R)-isopropyl 2-amino-3-hydroxybutanoate hydrochloride (169.9 mg,33%) as a white solid.

¹H NMR (400 MHz, CHLOROFORM-d) ⋅ 8.35 (br. s., 3H), 5.18 (td, J=6.24,12.47 Hz, 1H), 4.48-4.65 (m, 1H), 4.31 (m, 1H), 1.38 (d, J=6.60 Hz, 3H),1.33 (d, J=6.36 Hz, 6H).

Intermediate 290: Isopropyl 2-amino-3-hydroxybutanoate hydrochloride

Acetyl chloride (35.8 ml, 504 mmol) was added drop wise to propan-2-ol(240 ml, 3115 mmol) while cooling in an ice bath, then the mixture wasstirred for 30 min at room temperature. After that2-amino-3-hydroxybutanoic acid (20 g, 168 mmol) was added to thereaction mixture and then reaction mixture was heated at 80° C. for 16h.The reaction mixture concentrated under reduced pressure and theresultant residue was azeotroped with Toluene (2×100 mL), thentriturated with ether (100 mL) and dried to get the title compoundisopropyl 2-amino-3-hydroxybutanoate, hydrochloride (24 g, 121 mmol,72.3% yield) as a white semi solid. LCMS (System G): t_(RET)=1.39 min;MH⁺ 162.

Intermediate 291:5-(1-(1,3-dimethoxypropan-2-yl)-5-(oxiran-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde(for an example preparation see Intermediate 224, 2.0 g, 5.41 mmol),trimethyl sulfonium iodide (1.127 g, 5.52 mmol) and potassium hydroxide(1.823 g, 32.5 mmol) in acetonitrile (20 mL)/water (0.5 mL) were stirredunder nitrogen at room temperature. The reaction mixture was thenstirred at 65° C. for 2 h. The reaction was then quenched with water (20mL) and extracted with ethyl acetate (50 mL×2), the combined organiclayer was washed with brine, dried (Na₂SO₄), filtered and concentratedto give the title product5-(1-(1,3-dimethoxypropan-2-yl)-5-(oxiran-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(2.0 g, yield 92%). The crude product was carried through to the nextstep without further purification. LCMS (System H): t_(RET)=4.31 min;MH⁺384.

Intermediate 292:1,3-dimethyl-5-(5-(oxiran-2-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)pyridin-2(1H)-one

To a solution of2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(For a preparation see Intermediate 116, 1 g, 2.62 mmol),trimethylsulfonium iodide (0.536 g, 2.62 mmol) in Acetonitrile (15 mL)and water (0.075 mL) stirred under nitrogen at room temp was added neatpotassium hydroxide (0.884 g, 15.75 mmol) in one charge at rt. Thereaction mixture was stirred at 65° C. for 3 hr. Reaction was dilutedwith EtOAc (50 mL) and reaction mixture was passed through hydrophobicfrit and filtrate was concentrated under reduced pressure to produce acrude residue. The crude residue was diluted with saturated sodiumbicarbonate solution (50 mL) and EtOAc (100 mL). The aqueous layer wasseparated and re-extracted with EtOAc (2×200 mL). The combined organicphases were washed with water (50 mL) and brine solution (50 mL). Theorganic layer was dried over Na₂SO₄, filtered and the filtrate wasconcentrated to afford the crude title compound1,3-dimethyl-5-(5-(oxiran-2-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)pyridin-2(1H)-one(900 mg, 62.3% yield) as a light yellow solid. The crude product wascarried through to the next step without further purification. LCMS(System H): t_(RET)=3.28 min; MH⁺ 380.

Intermediate 77:(3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)methanol

A solution of (tetrahydro-2H-pyran-4-yl)methanamine (5.23 mL, 40.9mmol), (4-fluoro-3-nitrophenyl)methanol (3.5 g, 20.45 mmol) andN-ethyl-N-isopropylpropan-2-amine (17.86 mL, 102 mmol) intetrahydrofuran (THF) (10 mL) was degassed and heated under nitrogen at60° C. for 5 hours. The reaction mixture was partitioned between ethylacetate (150 mL) and saturated solution of sodium bicarbonate (150 mL).The organic layer was isolated and the aqueous fraction was re-extractedtwice with ethyl acetate (2×150 mL). The organic fractions werecombined, passed through a hydrophobic frit and concentrated underreduced pressure. The residue was dissolved in a minimum amount of ethylacetate and loaded onto a silica column (100 g, SPE). The product waseluted with a gradient of 0-60% of ethyl acetate in cyclohexane. Productcontaining fractions were combined and concentrated under reducedpressure to yield the title compound (4957 mg, 91% yield) as an orangesolid. LCMS (System B): t_(RET)=0.79 min; MH⁺ 267.

Intermediate 96:5-(5-(hydroxymethyl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one

To a solution of(3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)methanol (foran example preparation see Intermediate 77, 2.05 g, 7.70 mmol) inethanol (60 mL) was added1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (for an examplepreparation see Intermediate 2, 1.455 g, 7.70 mmol) and sodiumhydrosulfite (4.76 g, 23.09 mmol), followed by water. The reactionmixture was heated at 90° C. for 2 days. The reaction mixture wasconcentrated to approximately half the volume under reduced pressure,and the resulting liquid partitioned between 3:1 chloroform:isopropanol(3×150 mL) and saturated aqueous sodium bicarbonate solution (150 mL).The organic layers were combined, dried using a hydrophobic frit andevaporated under reduced pressure. The sample was loaded indichloromethane and purified by Biotage SP4 SNAP 100 g silica using agradient of 0-25% cyclohexane-(25% ethanol in ethyl acetate) over 10column volumes followed by holding at 25% cyclohexane-(25% ethanol inethyl acetate) for 5 column volumes, followed by a gradient of 0-100%cyclohexane-(25% ethanol in ethyl acetate) over 10 column volumes,followed by 25% ethanol in ethyl acetate over 10 column volumes. Theappropriate fractions were combined and evaporated under reducedpressure to give the required product5-(5-(hydroxymethyl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(1.22 g, 3.32 mmol, 43.1% yield) as an off-white foam. LCMS (System B):t_(RET)=0.63 min; MH⁺ 368.

Intermediate 116:2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

5-(5-(hydroxymethyl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for an example preparation see Intermediate 96, 7 g, 19.05 mmol) wasdissolved in dichloromethane (DCM) (200 mL) and manganese dioxide (6.62g, 76 mmol) was added, the mixture was heated at reflux for 2h beforethe mixture was filtered and the solid washed with DCM. The filtrate wasevaporated in vacuo to give2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(6.6 g, 18.06 mmol, 95% yield) as a pale yellow foam. LCMS (System J):t_(RET)=0.75 min; MH⁺ 366.

Example Preparation Example 1: (S)-cyclopentyl4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)pentanoate

A round bottom flask was charged with5-methyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (For an examplepreparation see Intermediate 1, 203 mg, 1.482 mmol), sodium hydrosulfite(821 mg, 4.72 mmol), Water (10 mL), and a solution of(S)-cyclopentyl-4-methyl-2-((3-nitro-4-(((tetrahydro-2Hpyran-4-yl)methyl)amino)benzyl)amino)pentanoate (For an examplepreparation, see Intermediate 182, 603 mg, 1.347 mmol) in Ethanol (20mL). A condenser was fitted and the slurry refluxed at 100° C.overnight. The mixture was diluted with water and EtOAc, the layersmixed and separated before the organic layer was passed through ahydrophobic frit and concentrated in vacuo to give a yellow oil. Thesample was loaded in dichloromethane and purified by silica gel columnchromatography using a gradient of 0-100% ethyl acetate-cyclohexane over15 column volumes followed 0-10% 2M ammonia in methanol-dichloromethanefor 15 column volumes. The appropriate fractions were combined andevaporated in vacuo to give the title compound (140 mg) as a colourlessoil that solidified under high vacuum. LCMS (System A): t_(RET)=0.78min; MH⁺=535.

The following Examples were prepared in a similar manner to Example 1:(In the tables, details of the LCMS system used, retention time(t_(R)ET), MH⁺, reaction yield and % yield are provided for eachExample).

Example 2: (S)-cyclopentyl 4- methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1- methylpiperidin-4-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino) pentanoate (prepared from: Intermediate181) System B, 1.10 min, MH⁺ = 534, Yield: 38 mg, 12%

Example 3: (S)-cyclopentyl 2- (((1-(2-methoxyethyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin- 3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-4-methylpen- tanoate (prepared from: Interme- diate 184)System A, 0.82 min, MH⁺ = 495, Yield: 206 mg, 48%

Example 4: (S)-cyclopentyl 2- (((1-(2-(dimethylamino)ethyl)-2-(5-methyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)- 4-methylpentanoate (prepared from:Intermediate 185) System A, 0.69 min, MH⁺ = 508, Yield: 240 mg, 37%

Example 5: (S)-cyclopentyl 2- (((1-(3-hydroxypropyl)-2-(5-methyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)- 4-methylpentanoate (prepared from:Intermediate 186) System A, 0.75 min, MH⁺ = 495, Yield: 120 mg, 22%

Example 6: (S)-cyclopentyl 4- methyl-2-(((1-methyl-2-(5-methyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino) pentanoate (prepared from: Intermediate 187)System A, 0.73 min, MH⁺ = 451, Yield: 35 mg, 33%

Example 7: (2S)-cyclopentyl 4- methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((4- methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)pentanoate (prepared from:Intermediate 143) System A, 0.65 min, MH⁺ = 550, Yield: 290 mg, 35%

Example 8: (2S)-cyclopentyl 2-(((2- (5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((4-methylmorpholin-2-yl) methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)propanoate (prepared from: Intermediate 145) System B, 0.89min, MH⁺ = 508, Yield: 119 mg, 15%

Example 9: (2S)-cyclopentyl 3-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1-((4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-5- yl)methyl)amino)butanoate (preparedfrom: Intermediate 147) System B, 1.06 min, MH⁺ = 536, Yield: 348 mg,38%

Example 10: (S)-cyclopentyl 4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydro-pyridin-3-yl)-1-((1-methylpiperidin-4-yl)methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)pentanoate (preparedfrom: Intermediate 149) System B, 1.14 min, MH⁺ = 548, Yield: 237 mg,34%

Example 11: (S)-tert-butyl 4-methyl- 2-(((2-(5-methyl-6-oxo-1,6-dihydro-pyridin-3-yl)-1-(1-methylpiperidin-4-yl)-1H-benzo[d]imidazol-5-yl)methyl) amino)pentanoate (prepared from:Intermediate 189) System B, 1.07 min, MH⁺ = 522, Yield: 36 mg, 25%

Example 12: (S)-cyclopentyl 4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydro-pyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-6- yl)methyl)amino)pentanoate (preparedfrom: Intermediate 190) System B, 1.14 min, MH⁺ = 535, Yield: 74 mg, 25%

Example 7a and 7b: (S)-cyclopentyl4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(((R)-4-methylmorpholin-2-yl)methyl)-1-benzo[d]imidazol-5-yl)methyl)amino)pentanoateand (S)-cyclopentyl4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(((S)-4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)pentanoate

Example 7 (95 mg) was dissolved in Ethanol (2 mL) and purified by chiralchromatography (stationary phase: Chiralpak AD-H 5 μm particles, mobilephase: heptane/ethanol) (N20771-72). The combined fractions containingpure Isomer 1 were concentrated in vacuo and the solid residue wastransferred to a weighed flask using a mixture of methanol anddichloromethane, which was removed in vacuo. The material was dried in avacuum oven at 45° C. for 4 hours and at ambient temperature for 24hours to give pure Isomer 1 (Example 7a) as a colourless solid (39.8mg). HPLC-UV: RT 16.73 minutes, ca. 99.7% isomeric purity by area HPLC @280 nM. LCMS (System B): t_(RET)=1.11 min, MH⁺=550. The combinedfractions containing pure Isomer 2 were concentrated in vacuo and thesolid residue was transferred to a weighed flask using a mixture ofmethanol and dichloromethane, which was removed in vacuo. The materialwas dried in a vacuum oven at 45° C. for 4 hours and at ambienttemperature for 24 hours to give pure Isomer 2 (Example 7b) of thebenzimidazole as a colourless solid (41.2 mg). HPLC-UV: RT 21.09minutes, ca. 99.4% isomeric purity by area HPLC @ 280 nM. LCMS (SystemB): t_(RET)=1.12 min, MH⁺=550.

Example 13: (2S)-cyclopentyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazole-5-yl)methyl)amino)-4-methylpentanoate

1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (For an examplepreparation see Intermediate 2, 196 mg, 1.297 mmol) and sodiumdithionite (677 mg, 3.89 mmol) were added to a solution of(2S)-cyclopentyl4-methyl-2-((4-(((4-methylmorpholin-2-yl)methyl)amino)-3-nitrobenzyl)amino)pentanoate(For a preparation see Intermediate 143, 600 mg, 1.297 mmol) in Ethanol(10 mL) and Water (5 mL) and the reaction mixture was heated bymicrowave irradiation to 100° C. for 5 hours. The reaction mixture waspartitioned between DCM (40 mL) and saturated sodium hydrogen carbonatesolution (40 mL) and the layers were separated. The aqueous layer wasextracted with DCM (3×40 mL) and the combined organic layers were driedusing a hydrophobic frit and evaporated under reduced pressure. Thecrude sample was purified by silica gel column chromatography using agradient of 0-10% dichloromethane-2M ammonia in methanol over 10 columnvolumes followed by 10% dichloromethane-2M ammonia in methanol for 5column volumes. The appropriate fractions were combined and evaporatedunder reduced pressure. This crude sample was purified twice by MDAP(Method B) The appropriate fractions were combined and the solvent wasevaporated under reduced pressure to give the title compound (202 mg) asan off-white solid. LCMS (System B): t_(RET)=1.16 min; MH⁺=564.

The following Examples were prepared in a similar manner to Example 13:

Example 14: (S)-cyclopentyl 2- (((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetra- hydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-4-methyl- pentanoate (preparedfrom: Intermediate 182) System A, 0.85 min, MH⁺ = 548, Yield 140 mg, 27%

Example 15: (S)-cyclopentyl 2- (((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-methyl- 1H-benzo[d]imidazol-5-yl)methyl)amino)-4-methyl- pentanoate (prepared from: Intermediate 187)System A, 0.77 min, MH⁺ = 465, Yield 33 mg, 30%

Example 16: (S)-cyclopentyl3-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1-benzo[d]imidazol-5-yl)methyl)amino)butanoate

A suspension of2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(200 mg, 0.569 mmol, Intermediate 151) and (S)-cyclopentyl2-amino-3-methylbutanoate 4-methylbenzenesulfonate (407 mg, 1.138 mmol,Intermediate 24) in DCM (10 mL) was stirred under nitrogen at roomtemperature. After 15 minutes, partial dissolution was achieved andsodium triacetoxyborohydride (241 mg, 1.138 mmol) was added to this. Thereaction mixture was stirred under nitrogen at room temperature for 2hours. The reaction mixture was partitioned between DCM (30 mL) andsaturated sodium hydrogen carbonate solution (30 mL) and the layers wereseparated. The aqueous layer was extracted with DCM (3×30 mL) and theorganic layers were combined, dried using a hydrophobic frit andevaporated under reduced pressure to give a white solid. The crudesample was dissolved in DMSO (3×1 mL) and purified by MDAP (Method B).The solvent was evaporated under reduced pressure to give the titlecompound (196.1 mg, 0.377 mmol, 66.2% yield) as a white solid. LCMS(System B): t_(RET)=1.11 min, MH⁺=521.

Example 17:(S)-4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)pentanoicacid, hydrochloride

To a solution of (S)-cyclopentyl4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)pentanoate(For a preparation see Example 12, 35 mg, 0.065 mmol) in Methanol (2 mL)and THF (2 mL) was added 1 M aqueous lithium hydroxide (0.196 mL, 0.196mmol), and the reaction mixture heated at 50° C. overnight. The reactionmixture was blown down under a stream of nitrogen. The sample wasdissolved in 2 M aqueous hydrochloric acid (0.1 mL) and methanol (0.9mL) and purified by MDAP (Method B). The solvent was blown down under astream of nitrogen to give a white gum. The sample was suspended intetrahydrofuran (1 mL) and 2 M aqueous hydrochloric acid (0.5 mL) addedto give a clear solution. The mixture was blown down under a stream ofnitrogen to give the title compound (22 mg) as an off-white solid. LCMS(System B): t_(RET)=0.55 min; MH⁺ 467.

The following Examples were prepared in a similar manner to Example 17:

Example 18: (S)-2-(((1-ethyl- 2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H- benzo[d]imidazol-5-yl) methyl)amino)-4-methyl-pentanoic acid, hydrochloride (prepared from: Example 151) System B,0.58 min, MH⁺ = 397, Yield 88 mg, 23%

Example 19: (S)-2-(((1-(2- methoxyethyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3- yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-4-methyl- pentanoic acid, hydrochloride (prepared from:Example 3) System A, 0.48 min, MH⁺ = 427, Yield: 29 mg, 89%

Example 20: (S)-2-(((1-(2- (dimethylamino)ethyl)-2-(5- methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H- benzo[d]imidazol-5-yl) methyl)amino)-4-methyl-pentanoic acid, hydrochloride (prepared from: Example 4) System A, 0.38min, MH⁺ = 440, Yield: 35 mg, 107%, (hygroscopic product)

Example 21: (S)-2-(((1-(3- hydroxypropyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3- yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-4-methyl- pentanoic acid, hydrochloride (prepared from:Example 5) System A, 0.42 min, MH⁺ = 427, Yield: 28 mg, 100%

Example 22: (S)-2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydro-pyridin-3-yl)-1-((tetrahydro- 2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl) amino)-4-methylpentanoic acid (preparedfrom: Example 14) System A, 0.52 min, MH⁺ = 481, Yield: 19 mg, 67%

Example 23: (S)-2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydro-pyridin-3-yl)-1-methyl-1H- benzo[d]imidazol-5-yl)methyl)amino)-4-methylpentanoic acid, hydrochloride (prepared from: Example 15)System A, 0.45 min, MH⁺ = 397, Yield: 8.7 mg, 93%

Example 24: (S)-4-methyl-2- (((1-methyl-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H- benzo[d]imidazol-5-yl)methyl)amino)pentanoic acid, hydro- chloride (prepared from: Example 6) SystemA, 0.42 min, MH⁺ = 383, Yield: 4.6 mg, 50%

Example 25: (S)-2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydro-pyridin-3-yl)-1-ethyl-1H- benzo[d]imidazol-5-yl)methyl)amino)-3-methoxypropanoic acid, hydrochloride (prepared from: Example195) System A, 0.43 min, MH⁺ = 399, Yield: 12 mg, 31%

Example 26: (S)-2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydro-pyridin-3-yl)-1-ethyl-1H- benzo[d]imidazol-5-yl) methyl)amino)-4-methyl-pentanoic acid, hydrochloride (prepared from: Example 198) System A,0.50 min, MH⁺ = 411, Yield: 21 mg, 54%

Example 27: (S)-2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydro-pyridin-3-yl)-1-(2-methoxy- ethyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3- methoxypropanoic acid, hydrochloride (preparedfrom: Example 201) System A, 0.45 min, MH⁺ = 429, Yield: 22 mg, 55%

Example 28: (2S,3R)-2-(((1- ethyl-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxy- butanoic acid, hydrochloride (prepared from:Example 95) System A, 0.38 min, MH⁺ = 385, Yield: 14 mg, 38%

Example 29: (2S,3R)-2-(((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-ethyl- 1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxy- butanoic acid, hydrochloride (prepared from:Example 199) System A, 0.40 min, MH⁺ = 399, Yield: 16 mg, 44%

Example 30: (2S,3R)-3- hydroxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin- 3-yl)-1-(oxetan-3-ylmethyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)butanoic acid hydrochloride(prepared from: Example 200) System A, 0.37 min, MH⁺ = 427, Yield: 11mg, 27%

Example 31: (2S,3R)-3- hydnoxy-2-(((1-(2- methoxyethyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin- 3-yl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)butanoic acid hydrochloride (prepared from: Example 197) System A, 0.39min, MH⁺ = 415, Yield: 16 mg, 41%

Example 32: (S)-2-(((2- (1,5-dimethyl-6-oxo- 1H-dihydropyridin-3-yl)-1-methyl-1H-benzo[d] imidazol-5-yl)methyl) amino)-3-methoxy- propanoicacid hydro- chloride (prepared from: Example 300) System A, 0.37 min,MH⁺ = 385, Yield: 7.1 mg, 20%

Example 33: (S)-2-(((2- (1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-methyl-1H-benzo[d] imidazol-5-yl)methyl) amino)-3-methylbutanoic acidhydrochloride (prepared from: Example 285) System A, 0.38 min, MH⁺ =383, Yield: 3.2 mg, 9%

Example 34: (2S,3R)-2- (((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-methyl-1H-benzo[d] imidazol-5-yl)methyl)amino)-3-hydroxy- butanoic acid hydro- chloride (prepared from: Example155) System A, 0.34 min, MH⁺ = 385, Yield: 1.7 mg, 5%

Example 35: (2S,3R)-2- (((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxyethyl)- 1H-benzo[d]imidazol-5- yl)methyl)amino)-3-hydroxybutanoic acid hydrochloride (prepared from: Example 204) SystemA, 0.39 min, MH⁺ = 429, Yield: 10 mg, 25%

Example 36: (2S,3R)-2- (((1-(2-(dimethylamino) ethyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin- 3-yl)-1H-benzo[d] imidazol-5-yl)methyl)amino)-3-hydroxy- butanoic acid dihydro- chloride (prepared from:Example 203) System A, 0.29 min, MH⁺ = 428, Yield: 6 mg, 13%

Example 37: (2S,3R)-2- (((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-l-((tetrahydro-2H- pyran-4-yl)methyl)-1H- benzo[d]imidazol-6-yl)methyl)amino)-3- hydroxybutanoic acid hydrochloride (prepared from:Example 105) System J, 0.52 min,

MH⁺ = 469, Yield: 58 mg, 62% Example 38: (2S,3R)-2-(((1-((1,4-dioxan-2-yl) methyl)-2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1H-benzo [d]imidazol-5-yl)methyl) amino)-3-hydroxy-butanoic acid hydro- chloride (prepared from: Example 109a) System B,0.49 min, MH⁻ = 469, Yield: 30 mg, 48%

Example 39: (2S,3R)-2- (((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)-1H- benzo[d]imidazol-5-yl)methyl)amino)-3- methoxybutanoic acid hydrochloride (prepared from:Example 114) System J, 0.54 min, MH⁺ = 469 Yield: 81 mg, 46%

Example 40: (2S,3R)-2- (((1-(((R)-1-acetyl- piperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo- 1,6-dihydropyridin-3- yl)-1H-benzo[d]imidazol-6-yl)methyl) amino)-3-hydroxy- butanoic acid hydro- chloride(prepared from: Example 124) System J, 0.52 min, MH⁺ = 510, Yield: 21mg, 77%

Example 41: (2S,3R)- 2-(((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-1-((S)-piperidin- 3-ylmethyl)-1H-benzo [d]imidazol-6-yl)methyl)amino)-3- hydroxybutanoic acid hydrochloride (prepared from:Example 124) System J, 0.49 min, MH⁺ = 468, Yield: 6 mg, 22%

Example 42a: (2S,3R)- 2-(((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxy- propyl)-1H-benzo[d] imidazol-5-yl)methyl)amino)-3-hydroxy- butanoic acid hydro- chloride (single diastereomer ofun- known configuration at marked position, Isomer 1) (prepared from:Example 271b) System

J, 0.55 min, MH⁺ = 443, Yield: 65 mg, 50% Example 42b: (2S,3R)-2-(((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin- 3-yl)-1-(2-methoxy-propyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)-3-hydroxy- butanoicacid hydro- chloride (single diastereomer of un- known configuration atmarked position, Isomer 2) (prepared from: Example 271a) System

J, 0.55 min, MH⁺ = 443, Yield: 50 mg, 38% Example 43: (2S,3R)-2-(((1-(((S)-1-acetyl- piperidin-3-yl)methyl)- 2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1H- benzo[d]imidazol-6-yl)methyl)amino)-3- hydroxybutanoic acid hydrochloride (prepared from:Example 128) System J, 0.51 min, MH⁺ = 510, Yield: 10 mg, 51%

Example 44: (2S,3R)-2- (((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxy- butan-2-yl)-1H-benzo [d]imidazol-5-yl)methyl)amino)-3-hydroxy- butanoic acid hydro- chloride (Single diastereomer ofun- known configuration at marked position) (prepared from: Example131a) System

I, 0.48 min, MH⁺ = 457, Yield: 13 mg, 67% Example 45: (2S)-4-methyl-2-(((2-(5- methyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((4-methylmorpholin-2- yl)methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino) pentanoic acid, bis- hydrochloride (prepared from: Example7) System B, 0.56 min, MH⁺ = 482, Yield: 50 mg, 50%

Example 46: (2S)-2- (((2-(5-methyl-6-oxo- 1,6-dihydropyridin-3-yl)-1-((4-methyl- morpholin-2-yl) methyl)-1H-benzo[d]imidazol-5-yl)methyl) amino)propanoic acid, bis hydrochloride (preparedfrom: Example 8) System B, 0.48 min, MH⁺ = 440, Yield: 32 mg, 57%

Example 47: (2S)-3- methyl-2-(((2-(5- methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((4-methylmorpholin- 2-yl)methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)butanoic acid, bis hydrochloride(prepared from: Example 9) System B, 0.52 min, MH⁺ = 468, Yield: 82 mg,82%

Example 48: (S)-4- methyl-2-(((2-(5-methyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1-((1- methylpiperidin-4-yl) methyl)-1H-benzo[d]imidazol-5-yl)methyl) amino)pentanoic acid, bis hydrochloride (preparedfrom: Example 10) System B, 0.57 min, MH⁺ = 480, Yield: 56 mg, 56%

Example 49: (2S)-2-(((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1- ((4-methylmorpholin-2- yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl) amino)-4-methyl- pentanoic acid, bis hydrochloride(prepared from: Example 13) System B, 0.58 min, MH⁺ = 496, Yield: 56 mg,55%

Example 50: (S)-3-methyl- 2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)butanoic acid,hydrochloride (prepared from: Example 16) System B, 0.53 min, MH⁺ = 453,Yield: 28 mg, 60%

Example 51: (S)-3-hydroxy- 2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1- ((tetrahydro-2H-pyran-4- yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl) amino)propanoic acid, hydrochloride 9preparedfrom: Example 137) System B, 0.48 min, MH⁺ = 441, Yield: 33 mg, 70%

Example 52: (2S,3R)-3- hydroxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin- 3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H- benzo[d]imidazol-5-yl) methyl)amino)butanoicacid, hydrochloride (prepared from: Example 138) System B, 0.49 min, MH⁺= 455, Yield: 40 mg, 86%

Example 53: (S)-2-(((2- (5-methyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino)propanoic acid, hydrochloride (prepared from: Example 140) SystemB, 0.49 min, MH⁺ = 425, Yield: 43 mg, 91%

Example 54: (S)-2-(((2- (5-methyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino)-2-(tetrahydro- 2H-pyran-4-yl)acetic acid, hydrochloride (preparedftrom: Example 142) System B, 0.51 min, MH⁺ = 495, Yield: 30 mg, 51%

Example 55: (S)-4- methoxy-2-(((2-(5- methyl-6-oxo-1,6-dihydropyridin-3-yl)-1- ((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)butanoic acid, hydrochloride (preparedfrom: Example 143) System B, 0.51 min, MH⁺ = 469, Yield: 34 mg, 65%

Example 56: (S)-2-(((2- (5-methyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino)butanoic acid, hydrochloride (prepared from: Example 144) SystemB, 0.50 min, MH⁺ = 439, Yield: 40 mg, 73%

Example 57: (S)-2-(((2- (5-methyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino)pentanoic acid, hydrochloride (prepared from: Example 145) SystemB, 0.54 min, MH⁺ = 453, Yield: 38 mg, 85%

Example 58: (S)-3- methoxy-2-(((2-(5- methyl-6-oxo-1,6-dihydropyridin-3-yl)-1- ((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl) amino)propanoic acid, hydrochloride (preparedfrom: Example 146) System B, 0.50 min, MH⁺ = 455, Yield: 34 mg, 72%

Example 59: (2S,3R)-2- (((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((S)-1-methoxypropan- 2-yl)-1H-benzo[d]imidazol-5-yl)methyl) amino)-3-hydroxy- butanoic acid, hydro- chloride(prepared from: Example 218) System A, 0.41 min, MH⁺ = 443, Yield: 65mg, 64%

Example 60: (S)-4-methyl- 2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(1-methylpiperidin-4-yl)-1H-benzo[d]imidazol-5- yl)methyl)amino)pentanoic acid, bis-hydrochloride(prepared from: Example 2) System B, 0.56 min, MH⁺ = 466, Yield: 26 mg,76%

Example 61: (S)-3-(1H- imidazol-5-yl)-2-(((2- (5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1- ((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl) amino)propanoic acid, bis hydrochloride(prepared from: Example 192) System B, 0.48 min, MH⁺ = 491, Yield: 26mg, 76%

Example 62: (2S,3R)-2- (((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1- (((S)-tetrahydrofuran-2-yl)methyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)-3-hydroxybutanoic acidhydrochloride (prepared from: Example 110) System J, 0.56 min, MH⁺ =455, Yield: 57 mg, 73%

Example 63: (2S,3R)-2- (((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-1-(((R)-tetrahydro- 2H-pyran-3-yl)methyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoic acid hydrochloride (prepared from:Example 213a) System J, 0.57 min, MH⁺ = 469, Yield: 16 mg, 48%

Example 64: (2S,3R)-2- (((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-1-(((S)-tetrahydro- 2H-pyran-3-yl)methyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoic acid hydrochloride (prepared from:Example 213b) System J, 0.57 min, MH⁺ = 469, Yield: 24 mg, 61%

Example 65: (S)-2-(((2- (1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino)-3-hydroxy- propanoic acid hydro- chloride (prepared from: Example233) System J, 0.52 min, MH⁺ = 455, Yield: 53 mg, 51%

Example 66: (2S,3R)-2- (((1-(1,3-dimethoxy- propan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoic acid hydrochloride (prepared from:Example 305) System J, 0.54 min, MH⁺ = 473, Yield: 19 mg, 96%

Example 67: (2S,3R)-2- ((2-((2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1-((tetra- hydro-2H-pyran-4-yl) methyl)-1H-benzo[d]imidazol-5-yl)oxy)ethyl) amino)-3-hydroxy- butanoic acid, Hydro-chloride (prepared from: Example 276) System J, 0.55 min, MH⁺ = 499,Yield: 59 mg, 35%

Example 68: (2S,3R)-2- (((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)-1H- benzo[d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoic acid hydrochloride (prepared from:Example 287) System C, 0.41 min, MH⁺ = 469, Yield: 40 mg, 41%

Example 69: (2S,3R)-2- (((1-(cyclopropylmethyl)- 2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1H-benzo[d]imidazol-6- yl)methyl)amino)-3-hydroxybutanoic acid hydrochloride (prepared from: A mixture of Example170 and Example 169) System I, 0.39 min, MH⁺ = 425,

Yield: 38 mg 78% Example 70a: (2S,3R)- 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin- 3-yl)-1-((tetrahydro-2H- pyran-3-yl)methyl)-1H-benzo[d]imidazol-6- yl)methyl)amino)-3- hydroxybutanoic acidhydrochloride (Single diastereomer of un- known configuration, Isomer 1)(prepared from: Example 171a)

System I, 0.40 mins, MH⁺ = 469, Yield: 22 mg 65% Example 70b: (2S,3R)-2-(((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin- 3-yl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H- benzo[d]imidazol-6- yl)methyl)amino)-3-hydroxybutanoic acid hydrochloride (Single diastereomer of un- knownconfiguration, Isomer 2) (prepared from: Example 171b)

System I, 0.41 mins, MH⁺ = 469, Yield: 16 mg 70%

Example 71:(S)-3,3-dimethyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoicacid

1M aq Lithium hydroxide solution (1.0 mL) was added to a solution of(2S)-tetrahydrofuran-3-yl3,3-dimethyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoate(For a preparation see Example 239, 50 mg, 0.093 mmol) in methanol (0.5mL) and THF (0.5 mL). The reaction mixture was stirred at 60° C. for 48hours. The reaction mixture was cooled to room temperature. The solventwas evaporated and the residue purified by MDAP (Method B) to give thetitle compound (26 mg), as a colourless solid. LCMS (System B):t_(RET)=0.57 min; MH⁺ 467 The following Examples were prepared in asimilar manner to Example 71:

Example 72: (2S)-2- (((1-((1-acetyl- pyrrolidin-3-yl)methyl)-2-(5-methyl- 6-oxo-1,6-dihydro- pyridin-3-yl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 4-methylpentanoic acid (preparedfrom: Example 153) System B, 0.53 min, MH⁺ = 494, Yield: 24 mg 97%

Example 73: (2S,3R)- 2-(((2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1-((S)-1- methoxypropan-2-yl)- 1H-benzo[d]imidazol-6-yl)methyl)amino)- 3-hydroxybutanoic acid (prepared from: Example 116)System J, 0.50 min, MH⁺ =

443, Yield: 61 mg 66% Example 74: (2S,3R)- 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1-((4- methylmorpholin-2-yl)methyl)-1H-benzo [d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoicacid (prepared from: Exam- ple 162) System B, 0.51 min, MH⁺ = 484,Yield: 3.5 mg, 20%

Example 75: (2S,3R)- 2-(((2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1-((4- methylmorpholin-3- yl)methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoic acid (prepared from:Example 165) System B, 0.46 min, MH⁺ = 484, Yield: 23 mg, 92%

Example 76: (S)-3- hydroxy-3-methyl-2- (((2-(5-methyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoic acid (prepared from:Example 240) System B, 0.73 min, MH⁺ = 539, Yield: 16 mg 74%

Example 77: (S)-2- cyclopropyl-2-(((2- (5-methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) acetic acid (prepared from:Example 245) System B, 0.52 min, MH⁺ = 451, Yield: 18 mg 83%

Example 78: (S)-3- cyclopropyl-2-(((2- (5-methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) propanoic acid (prepared from:Exam- ple 246) System B, 0.54 min, MH⁺ = 465, Yield: 16 mg 73%

Example 79: (R)-3- cyclopropyl-2-(((2-(5- methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) propanoic acid (pre- pared from:Example 247) System B, 0.55 min, MH⁺ = 465, Yield: 15 mg 69%

Example 80: (2S,3R)- 2-(((2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1- isopropyl-1H-benzo [d]imidazol-5-yl) methyl)amino)-3-hydroxybutanoic acid (prepared from: Exam- ple 97) System B, 0.40 min,MH⁺ = 413, Yield: 27 mg 63%

Example 81: (2S)-2- (((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-1-((4-methyl- morpholin-3-yl) methyl)-1H-benzo[d]imidazol-5-yl)methyl) amino)-3-methoxy- propanoic acid (pre- pared from:Example 166) System B, 0.50 min, MH⁺ = 484, Yield: 14 mg 52%

Example 82: (2S,3R)- 2-(((2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1- ((tetrahydro-2H-pyran- 2-yl)methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoic acid (prepared from:Exam- ple 248) System B, 0.60 min, MH⁺ = 469, Yield: 15 mg 66%

Example 83: (S)-2-((2- ((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro- 2H-pyran-4-yl)methyl)- 1H-benzo[d]imidazol-6-yl)oxy)ethyl)amino)- 3-methoxypropanoic acid (prepared from: Example277) System J, 0.55 min, MH⁺ = 499, Yield: 12 mg 68%

Example 84: (S)-2-((2- ((2-(1,5-dimethyl-6- oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro- 2H-pyran-4-yl)methyl)- 1H-benzo[d]imidazol-6-yl)oxy)ethyl)amino) propanoic acid (pre- pared from: Example 278)System J, 0.54 min, MH⁺ = 469, Yield: 14 mg 80%

Example 85: (S)-2-((2- ((2-(1,5-dimethyl-6- oxo-1,6-dihydro-pyridin-3-yl)-1-((tetra- hydro-2H-pyran-4- yl)methyl)-1H-benzo[d]imidazol-6-yl)oxy) ethyl)amino)-3-methyl- butanoic acid (Preparedfrom: Example 280) System J, 0.59 min, MH⁺ = 497, Yield: 9 mg 52%

Example 86:(2S,3S)-2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoicacid

To a stirred solution of (2S,3S)-(S)-tetrahydrofuran-3-yl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(50.0 mg, 0.093 mmol) (for an example preparation see Example 306) inethanol (3 mL) was added 1 M lithium hydroxide (0.464 mL, 0.464 mmol).The resulting solution was stirred overnight at 40° C. 2 M HCl (0.300mL, 0.600 mmol) was added to the reaction mixture and stirred for 30min. The reaction mixture was loaded onto a SCX Cartridge, eluted withMeOH (15 CV) and followed by 2 M NH₃ in MeOH (15 CV). The basic fractionwas concentrated and evaporated in vacuo to give the title compound as awhite solid. The total yield of the reaction was 100%. LCMS (System C):t_(RET)=0.40 min, MH⁺=469.

The following Examples were prepared in a similar manner to Example 86:

Example 87: (2S,3R)- 2-(((2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1-(tetra- hydro-2H-pyran-4-yl)- 1H-benzo[d]imidazol-6-yl)methyl)amino)- 3-hydroxybutanoic acid (prepared from: Example 173)System J, 0.49 min, MH⁺ = 455, Yield: 18 mg 88%

Example 88: (2S,3R)- 2-(((2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1-((tetra- hydro-2H-pyran-2- yl)methyl)-1H-benzo[d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoic acid- diastereomer 2(pre- pared from: Interme- diate 174b) System I, 0.46 min, MH⁺ = 469,

Yield: 28 mg 96% Example 89: (2S,3R)- 2-(((1-(((S)-1-acetyl-piperidin-3-yl)methyl)- 2-(1,5-dimethyl-6-oxo- 1,6-dihydropyridin-3-yl)-1H-benzo[d] imidazol-5-yl)methyl) amino)-3-hydroxy- butanoic acid(prepared from: Intermediate 89) System C, 0.38 min, MH⁺ = 510, Yield:8.7 mg 38%

Example 90:(S)-4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)pentanoicacid, sodium salt

A round bottom flask was charged with (S)-cyclopentyl4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1h-benzo[d]imidazol-5-yl)methyl)amino)pentanoate (136 mg, 0.254 mmol,Example 1), tetrahydrofuran (THF) (2 mL), methanol (2 mL), water (1 mL)and lithium hydroxide (15 mg, 0.626 mmol). An air condenser was fittedand the mixture warmed to 50° C. overnight. Lithium hydroxide (15 mg,0.626 mmol) was added and the mixture heated to 50° C. overnight. Themixture was cooled to room temperature before being concentrated invacuo to give a white solid. The solid was dissolved in 1:1 MeOH:DMSO2×1 mL and purified by MDAP (Method A). The relevant fractions werecombined and the solvent was evaporated in vacuo to a white solid (86mg, 0.184 mmol). The solid was slurried in a DCM/MeOH/THF solution (9 mL1:1:1) prior to the addition of sodium hydroxide (93 μL, 0.186 mmol).The resultant solution was stirred for 5 minutes prior to removal of thevolatiles in vacuo to give the title compound (96 mg, 0.19 mmol, 77%yield) as a white solid. LCMS (System A): t_(RET)=0.49 min; MH⁺467.

Example 91:(2S,3R)-2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoicacid—Diastereomer 1, Hydrochloride

To a stirred solution of (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate—diastereomer1 (174a) (31.5 mg, 0.062 mmol) in Ethanol (2 mL) was added 1M lithiumhydroxide (0.308 mL, 0.308 mmol). The resulting reaction mixture wasstirred for 2 days at 40° C. The reaction mixture was loaded onto aBiotage SCX Cartridge, eluted with MeOH (15 CV) and 2M methanolicammonia (15 CV). The basic fraction was evaporated in vacuo to obtain awhite solid. To a suspension of the residue in Et₂O (0.5 mL) was added0.5M HCl in Et2O (0.1 mL). The suspension was stirred for 2 h, andevaporated in vacuo to give the title compound as a white solid. Thetotal yield of the reaction was 72%. LCMS (System I): t_(RET)=0.46 min,MH⁺=469.

Example 92: (2S,3R)-tert-butyl2-(((1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate

Example 93:(2S,3R)-2-(((1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoicacid

(2S,3R)-tert-butyl 2-amino-3-hydroxybutanoate hydrochloride (115 mg,0.541 mmol) was added to a stirred solution of1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-6-carbaldehyde(134) (100 mg, 0.271 mmol) in DCM (5 mL) under a nitrogen atmosphere for22 h. Sodium triacetoxyborohydride (172 mg, 0.812 mmol) was added, theresulting suspension stirred for 1 h. MeOH (5 mL) was added, thesolution stirred for 5 min and loaded on to a 5 g SCX cartridge. Thecartridge was eluted with MeOH (25 mL), followed by 2M methanolicammonia (25 mL). The basic fractions were evaporated in vacuo to a brownoil and purified by MDAP (method B). 2 M aq. HCl was added (0.5 mL) andthe product containing fractions were evaporated in vacuo, azeotropingwith EtOH to give a white solid. The residue was dissolved in DMSO:MeOH(1:1, 0.9 mL) and purified by MDAP. The product containing fractionswere evaporated to dryness, azeotroping with EtOH and PhMe to give thetitle compounds as white solids. The total yield of Example 92 was 25%.LCMS (System C): t_(RET)=0.57 min, MH⁺=529. The total yield of Example93 was 5%. LCMS (System C): t_(RET)=0.40 min, MH⁺=473.

Example 94a:(2S,3R)-2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-benzo[d]imidazole-5-yl)methyl)amino)-3-hydroxybutanoicacid, Hydrochloride—Diastereomer 1

(2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(182a) (12.5 mg, 0.023 mmol) was dissolved in 2M aq. HCl (2 mL, 4.00mmol). The reaction mixture was heated to 40° C. for 4 days. Thereaction mixture was evaporated in vacuo to give the title compound as awhite solid. Total yield of the reaction was 58%. LCMS (System C):t_(RET)=0.40 min, MH⁺=483.

Example 94b:(2S,3R)-2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-benzo[d]imidazole-5-yl)methyl)amino)-3-hydroxybutanoicacid, Hydrochloride—Diastereomer 2

(2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(182b) (12.5 mg, 0.023 mmol) was dissolved in 2M aq. HCl (2 mL, 4.00mmol). The reaction mixture was heated to 40° C. for 4 days. Thereaction mixture was evaporated in vacuo to give the title compound as awhite solid. Total yield of the reaction was 80%. LCMS (System C):t_(RET)=0.40 min, MH⁺=483.

Example 95: (2S,3R)-cyclopentyl2-(((1-ethyl-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate

To a mixture of1-ethyl-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde(For a preparation, see Intermediate 198, 134 mg, 0.476 mmol) in DCM (5mL) was added (2S,3R)-cyclopentyl 2-amino-3-hydroxybutanoate4-methylbenzenesulfonate (For a preparation see Intermediate 31,N27467-41-1, 342 mg, 0.953 mmol) and triethylamine (0.166 mL, 1.191mmol), and the reaction mixture stirred at room temperature for 15 mins.Sodium triacetoxyborohydride (202 mg, 0.953 mmol) was added, and thereaction mixture stirred at room temperature for 2.5 hours. Saturatedsodium bicarbonate solution (10 mL) was added and the organic layerseparated. The aqueous layer was extracted with further dichloromethane(10 mL), and the organic layers were combined, dried using a hydrophobicfrit and blown down under a stream of nitrogen. The sample was dissolvedin DMSO (2×1 mL) and purified twice by MDAP (Method B). The solvent wasblown down under a stream of nitrogen to give the title compound (126mg) as an off-white solid. LCMS (System B): t_(RET)=0.89 min; MH⁺ 453The following Examples were prepared in a similar manner to Example 95:

Example 96: (S)-tetra- hydro-2H-pyran-4-yl 2-(((1-(2-methoxy-ethyl)-2-(5-methyl-6- oxo-1,6-dihydro- pyridin-3-yl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-4- methylpentanoate (pre- pared from:Interme- diate 201 and Interme- diate 14) System B, 0.96 min, MH⁺ = 511,Yield: 113 mg, 61%

Example 97: (2S,3R)- neopentyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-isopropyl-1H-benzo [d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoate (prepared from: Inter- mediate 274 andInter- mediate 10) System B,

1.04 min, MH⁺ = 483, Yield: 90 mg, 43% Example 98: (2S,3R)- isobutyl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-isopropyl-1H-benzo [d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoate(prepared from: Inter- mediate 274 and Inter- mediate 11) System B,

0.98 min, MH⁺ = 469, Yield: 31 mg, 20% Example 99: (S)-cyclo- pentyl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-isopropyl-1H-benzo [d]imidazol-5-yl) methyl)amino)-3-methoxypropanoate (prepared from: Inter- mediate 274 and Inter- mediate196) System

B, 1.03 min, MH⁺ = 481, Yield: 110 mg, 67% Example 100: (2S,3R)-isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-(((S)-tetrahydrofuran- 2-yl)methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoate (prepared from: Inter- mediate 118 andInter- mediate 11) System B, 0.96 min, MH⁺ = 511, Yield: 36 mg, 22%

Example 101: (2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(((R)-tetrahydrofuran- 2-yl)methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 119 and Inter- mediate 11) System B, 0.96 min, MH⁺ = 511,Yield: 29 mg, 36%

Example 102: (2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imi- dazol-6-yl)methyl) amino)-3-hydroxy- butanoate (preparedfrom: Intermediate 121

and Intermediate 11) System B, 0.89 min, MH⁺ = 525, Yield: 170 mg, 28%Example 103: (2S,3R)- (S)-tetrahydrofuran-3- yl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1-((tetra- hydro-2H-pyran-4-yl)methyl)-1H-benzo[d] imidazol-6-yl)methyl) amino)-3-hydroxy- butanoate(prepared from: Intermediate 121

and Intermediate 40) System A, 0.48 min, MH⁺ = 539, Yield: 40 mg, 11%Example 104: (2S,3R)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1- ((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo[d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 121 and Inter-

mediate 31) System B, 0.82 min, MH⁺ = 511, Yield: 145 mg, 52% Example105: (2S,3R)- cyclopentyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1- ((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo[d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 121 and Inter-

mediate 137) System B, 0.89 min, MH⁺ = 537, Yield: 197 mg, 67% Example106: (2S,3R)- tert-butyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1- ((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo[d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 121) System

B, 0.88 min, MH⁺ = 525, Yield 207 mg, 72% Example 107: (2S,3R)-neopentyl 2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo [d]imidazol-6-yl)methyl)amino)-3- hydroxybutanoate (prepared from: Inter- mediate 121 andInter- mediate 10) System B, 0.96 min, MH⁺ = 539, Yield: 110 mg, 75%

Example 108: (2S,3R)- isopropyl 3-hydroxy-2- (((2-(5-methyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((tetrahydro-2H- pyran-3-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoate (prepared from:Intermediate 211 and Intermediate 31) System B, 0.82 min, MH⁺ = 497,Yield: 136 mg, 43%

Example 109a: (2S,3R)- isobutyl 2-(((1-((1,4- dioxan-2-yl)methyl)-2-(1,5-dimethyl-6-oxo- 1,6-dihydropyridin-3- yl)-1H-benzo[d]imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate (single diastereomer,of un- known configuration at dioxane chiral centre, Isomer 1) (pre-

pared from: Interme- diate 120 and Inter- mediate 11) System B, 0.88min, MH⁺ = 527, Yield: 158 mg, 65% Example 109b: (2S,3R)- isopropyl2-(((1-((1,4- dioxan-2-yl)methyl)-2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin-3- yl)-1H-benzo[d] imidazol-5-yl)methyl)amino)-3-hydroxy- butanoate (single diastereomer, of un- knownconfiguration at dioxane chiral centre, isomer 2) (pre-

pared from: Interme- diate 120 and Inter- mediate 31) System B, 0.80min, MH⁺ = 513, Yield: 83 mg, 75% Example 110: (2S,3R)- isopropyl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-(((S)-tetrahydrofuran- 2-yl)methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoate (prepared from: Inter- mediate 118 andInter- mediate 31) System B, 0.87 min, MH⁺ = 497, Yield: 7 mg, 5.2%

Example 111: (2S,3R)- tert-butyl 2-(((1-((1,4- dioxan-2-yl)methyl)-2-(1,5-dimethyl-6-oxo- 1,6-dihydropyridin-3- yl)-1H-benzo[d]imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate (single diastereomer,of un- known configuration at dioxane chiral centre) (prepared

from: Intermediate 120 and (2S,3R)-tert- butyl 2-amino-3-hydroxybutanoate hydrochloride (com- mercially available)) System B,0.87 min, MH⁺ = 527, Yield: 21 mg, 73% Example 112: (2S,3R)-2-hydroxy-2-methyl- propyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 6-yl)methyl)amino)- 3-hydroxybutanoate (ptreparedfrom: Inter- mediate 121 and Inter- mediate 7) System J, 0.74 min, MH⁺ =541, Yield: 35 mg, 47%

Example 113: (2S,3R)- tert-butyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(((S)-tetrahydro- furan-2-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxybutanoate (preparedfrom: Inter- mediate 118 and (2S,3R)-tert-butyl 2- amino-3-hydroxy-butanoate hydro- chloride (commer- cially available)) System J, 0.99min, MH⁺ = 511, Yield:

89 mg, 61% Example 114: (2S,3R)- (S)-tetrahydrofuran-3- yl2-(((2-(1,5-dimethyl- 6-oxo-1,6-dihydro- pyridin-3-yl)-1-((tetra-hydro-2H-pyran-4-yl) methyl)-1H-benzo[d] imidazol-5-yl)methyl)amino)-3-methoxy- butanoate (prepared from: Intermediate 116 andIntermediate 9) System J, 0.84 min, MH⁺ = 553, Yield: 218 mg, 72%

Example 115: (2S,3R)- (S)-tetrahydrofuran-3- yl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1-(((S)- tetrahydrofuran-2-yl)methyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)-3-methoxy- butanoate(prepared from: Intermediate 118 and Intermediate 9) System J, 0.88 min,MH⁺ = 539, Yield: 189 mg, 49%

Example 116: (2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1- ((S)-1-methoxypropan- 2-yl)-1H-benzo[d]imidazol-6-yl)methyl) amino)-3-hydroxy- butanoate (prepared from:Intermediate 123 and Intermediate

11) System J, 0.96 min, MH⁺ = 499, Yield: 324 mg, 60% Example 117:(2S,3R)- tert-butyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1- (((S)-tetrahydrofuran- 2-yl)methyl)-1H-benzo[d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 124 and (2S,3R)-tert-butyl 2-

amino-3-hydroxy- butanoate hydro- chloride (commer- cially available))System J, 0.54 min, MH⁺ = 511, Yield: 22 mg, 15% Example 118: (2S,3R)-isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-(((S)-tetrahydro- furan-2-yl)methyl)- 1H-benzo[d]imidazol-6-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate124 and Inter- mediate 11) System I,

0.56 min, MH⁺ = 511, Yield: 55 mg, 38% Example 119: (2S,3R)- isopropyl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-(((S)-tetrahydro- furan-2-yl)methyl)- 1H-benzo[d]imidazol-6-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate124 and Inter-

mediate 31) System I, 0.51 min, MH⁺ = 497, Yield: 49 mg, 34% Example120: (2S,3R)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((S)-1-methoxy- propan-2-yl)-1H-benzo[d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 123 and Inter-

mediate 31) System J, 0.88 min, MH⁺ = 485, Yield: 41 mg, 35% Example123: (2S,3R)- tert-butyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((S)-1-methoxy- propan-2-yl)-1H-benzo[d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 123 and

(2S,3R)-tert-butyl 2- amino-3-hydroxy- butanoate hydro- chloride(commer- cially available)) System J, 0.95 min, MH⁺ = 499, Yield: 66 mg,56% Example 124: (2S,3R)- isopropyl 2-(((1-(((R)-1-acetylpiperidin-3-yl) methyl)-2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1H-benzo[d]imidazol- 6-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Inter- mediate 125 and Inter- mediate31) System J, 0.83 min, MH⁺ = 552, Yield: 85 mg, 41%

Example 125: (2S,3R)- isobutyl 2-(((1-(((R)-1- acetylpiperidin-3-yl)methyl)-2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1H-benzo[d]imidazol- 6-yl)methyl)amino)- 3-hydroxybutanoate (preparedfrom: Inter- mediate 125 and Inter- mediate 11) System J, 0.90 min, MH⁺= 566, Yield: 52 mg, 47%

Example 126: (2S,3R)- tert-butyl 2-(((1-(((R)- 1-acetylpiperidin-3-yl)methyl)-2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1H-benzo[d]imidazol- 6-yl)methyl)amino)- 3-hydroxybutanoate #(preparedfrom: Inter- mediate 125 and (2S,3R)-tert-butyl 2- amino-3-hydroxy-butanoate hydro- chloride (commer- cially available))

System J, 0.88 min, MH⁺ = 566, Yield: 58 mg, 83% Example 127: (2S,3R)-cyclobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((S)-1-methoxypropan- 2-yl)-1H-benzo[d] imidazol-6-yl)methyl)amino)-3-hydroxy- butanoate (prepared from: Intermediate 123 andIntermediate

32) System J, 0.93 min, MH⁺ = 497, Yield: 40 mg, 55% Example 128:(2S,3R)- isopropyl 2-(((1-(((S)- 1-acetylpiperidin-3-yl) methyl)-2-(1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1H-benzo[d]imidazol-6-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate127 and Inter- mediate 31) System J, 0.83 min, MH⁺ = 552, Yield: 87 mg,53%

Example 129: (2S,3R)- tert-butyl 2-(((1-(((S)- 1-acetylpiperidin-3-yl)methyl)-2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1H-benzo[d]imidazol- 6-yl)methyl)amino)- 3-hydroxybutanoate (preparedfrom: Inter- mediate 127 and (2S,3R)-tert-butyl 2- amino-3-hydroxy-butanoate hydro- chloride (commer-

cially available)) System B, 0.88 min, MH⁺ = 566, Yield: 19 mg, 34%Example 130: (2S,3R)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(1-methoxybutan- 2-yl)-1H-benzo[d]imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate (prepared from:Intermediate 117 and Intermediate 31) System I, 0.61 min, MH⁺ = 499,Yield: 278 mg, 66%

Example 131: (2S,3R)- cyclobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1- (1-methoxybutan-2-yl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoate (prepared from: Inter- mediate117 and Inter- mediate 32) System I, 0.62 min, MH⁺ = 511, Yield: 297 mg,69%

Example 132: (2S,3R)- tert-butyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1- (1-methoxybutan-2-yl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoate (prepared from: Inter- mediate117 and (2S,3R)-tert-butyl 2-amino-3-hydroxy- butanoate hydro- chloride(commer-

cially available)) System I, 0.63 min, MH⁺ = 513, Yield: 55 mg, 58%Example 133: (2S,3R)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydrofuran-3- yl)methyl)-1H-benzo[d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 129 and Inter-

mediate 31) System I, 0.84 min, MH⁺ = 497, Yield: 338 mg, 60% Example134: (2S,3R)- tert-butyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydrofuran-3- yl)methyl)-1H-benzo[d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 129 and

(2S,3R)-tert-butyl 2- amino-3-hydroxy- butanoate hydro- chloride(commer- cially available)) System J, 0.91 min, MH⁺ = 511, Yield: 224mg, 77% Example 135: (2S,3R)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1- ((S)-1-hydroxypropan- 2-yl)-1H-benzo[d]imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate (prepared from:Intermediate 236 and Intermediate 31) System I, 0.48 min, MH⁺ = 471,Yield:

67 mg, 47% Example 136: (S)-tert- butyl 4-methyl-2-(((2-(5-methyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-((tetrahydro-2H-pyran-4-yl)methyl)- 1H-benzo[d] imidazol-5-yl)methyl) amino)pentanoate(prepared from: Inter- mediate 151 and Inter- mediate X) System B, 1.14min, MH⁺ = 523, Yield: 18 mg, 88%

Example 137: (S)-cyclo- pentyl 3-hydroxy-2-(((2- (5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1- ((tetrahydro-2H-pyran- 4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl) amino)propanoate (pre- pared from: Interme-diate 151 and Interme- diate 276) System B, 0.81 min, MH⁺ = 509, Yield:234 mg, 81%

Example 138: (2S,3R)- cyclopentyl 3-hydroxy- 2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoate (prepared from:Intermediate 151 and Intermediate 137) System B, 0.87 min, MH⁺ = 523,Yield: 261 mg, 88%

Example 139: (2S)- tetrahydrofuran-3-yl 4-methyl-2-(((2-(5-methyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-((tetrahydro-2H-pyran-4-yl)methyl)- 1H-benzo[d] imidazol-5-yl)methyl) amino)pentanoate(Prepared from: Inter- mediate 151 and Inter- mediate 17) System B, 0.93min, MH⁺ = 537, Yield: 243 mg, 80%

Example 140: (S)- cyclopentyl 2-(((2-(5- methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)- 1H-benzo[d]imidazol-5-yl)methyl) amino)propanoate (prepared from: Inter- mediate151 and Inter- mediate 277) System B, 0.94 min, MH⁺ = 493, Yield: 176mg,

63% Example 141: (S)- neopentyl 4-methyl-2- (((2-(5-methyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((tetrahydro- 2H-pyran-4-yl)methyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)pentanoate (preparedfrom: Inter- mediate 151 and Inter- mediate 19) System B, 1.20 min, MH⁺= 537, Yield: 209 mg, 69%

Example 142: (S)- cyclopentyl 2-(((2- (5-methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)- 1H-benzo[d]imidazol-5-yl) methyl)amino)-2- (tetrahydro-2H-pyran- 4-yl)acetate(prepared from: Intermediate 151 and Intermediate 22) System B, 0.98min, MH⁺ = 563, Yield: 122 mg, 64%

Example 143: (S)- cyclopentyl 4- methoxy-2-(((2-(5- methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)- 1H-benzo[d]imidazol-5-yl) methyl)amino) butanoate (prepared from: Intermediate 151and Intermediate 23) System B, 0.96 min, MH⁺ = 537, Yield: 96 mg, 52%

Example 144: (S)- cyclopentyl 2-(((2- (5-methyl-6-oxo-1,6-dihydropyridin- 3-yl)-1-((tetrahydro- 2H-pyran-4-yl)methyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)butanoate (preparedfrom: Inter- mediate 151 and Inter- mediate 13) System B, 1.02 min, MH⁺= 507, Yield: 112 mg, 60%

Example 145: (S)- cyclopentyl 2-(((2-(5- methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) pentanoate (prepared from:Intermediate 151 and Intermediate 281) System B, 1.09 min, MH⁺ = 521,Yield 112 mg, 60%

Example 146: (S)- cyclopentyl 3-methoxy- 2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) propanoate (prepared from:Intermediate 151 and Intermediate 196) System B, 0.92 min, MH⁺ = 522,Yield: 127 mg, 71%

Example 147: (S)-(S)- 1-methoxypropan-2-yl 3-methyl-2-(((2-(5-methyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-((tetrahydro-2H-pyran-4-yl)methyl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoate(prepared from: Intermediate 151 and Intermediate 63) System A, 0.63min, MH⁺ = 525,

Yield: 52 mg, 28% Example 148: (S)-(R)- 1-methoxypropan-2-yl3-methyl-2-(((2-(5- methyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino) butanoate (prepared from: Intermediate 151 andIntermediate 280) System A, 0.62 min, MH⁺ = 525,

Yield: 103 mg, 57% Example 149: (2S,3R)- isobutyl 3-hydroxy-2-(((2-(5-methyl-6-oxo- 1,6-dihydropyridin-3- yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoate(prepared from: Intermediate 151 and Intermediate 11) System A, 0.87min, MH⁺ = 511, Yield: 72 mg, 28%

Example 150: (2S)- tert-butyl 4-methyl-2- (((2-(5-methyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((4-methyl- morpholin-2-yl)methyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)pentanoate (preparedfrom: Inter- mediate 275 and (S)- tert-butyl 2-amino-4- methylpentanoatehydrochloride (com- mercially available)) System B, 1.09 min,

MH⁺ = 538, Yield: 3.1 mg, 4% Example 151: (S)- cyclopentyl 2-(((1-ethyl-2-(5-methyl-6- oxo-1,6-dihydro- pyridin-3-yl)-1H-benzo[d]imidazol-5- yl)methyl)amino)-4- methylpentanoate (prepared from:Inter- mediate 113 and Inter- mediate 3) System B, 1.16 min, MH⁺ = 465,Yield: 65 mg, 33%

Example 152: (2S)- cyclopentyl 4-methyl- 2-(((1-((1-methyl-5-oxopyrrolidin-3-yl) methyl)-2-(5-methyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1H- benzo[d]imidazol-5- yl)methyl)amino) pentanoate(prepared from: Intermediate 153 and Intermediate 3) System B, 1.03 min,MH⁺ = 548, Yield: 23 mg, 13%

Example 153: (2S)- cyclopentyl 2-(((1- ((1-acetylpyrrolidin-3-yl)methyl)-2-(5- methyl-6-oxo-1,6- dihydropyridin-3-yl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 4-methylpentanoate (preparedfrom: Inter- mediate 156 and Inter- mediate 3) System B, 1.04 min, MH⁺ =562, Yield: 74 mg, 38%

Example 154: (2S,3R)- neopentyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-methyl-1H-benzo[d] imidazol-5-yl)methyl)amino)-3-hydroxy- butanoate (prepared from: Intermediate 273 andIntermediate

10) System B, 0.95 min, MH⁺ = 455, Yield: 137 mg, 85% Example 155:(2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-methyl-1H-benzo[d] imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate(prepared from: Intermediate 273 and Intermediate

11) System B, 0.89 min, MH⁺ = 441, Yield: 137 mg, 85% Example 156:(2S,3R)- cyclopentyl 2-(((2- (1,5-dimethyl-6-oxo- 1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate116 and Inter- mediate 137) System B, 0.91 min, MH⁺ = 537, Yield: 119mg, 75%

Example 157: (2S,3R)- (S)-tetrahydrofuran-3- yl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1- isopropyl-1H-benzo[d]imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate (prepared from:Intermediate 274 and Intermediate 40)

System A, 0.47 min, MH⁺ = 483, Yield: 111 mg, 71% Example 158: (2S,3R)-cyclopentyl 2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-isopropyl-1H- benzo[d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoate(prepared from: Inter- mediate 274 and Inter- mediate 137) System

A, 0.60 min, MH⁺ = 481, Yield: 83 mg, 53% Example 159: (2S,3R)-(S)-tetrahydrofuran-3- yl 2-(((2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1- (tetrahydro-2H-pyran- 4-yl)-1H-benzo[d]imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate (prepared from:Intermediate 161 and Intermediate 40) System A, 0.48 min, MH⁺ = 525,Yield: 33

mg, 25% Example 160: (2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(tetrahydro-2H- pyran-4-yl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 161 and Inter- mediate 11) System B, 0.89 min, MH⁺ = 511,Yield: 35 mg, 27%

Example 161: (2S,3R)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(tetrahydro-2H- pyran-4-yl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 161 and Inter- mediate 31) System I, 0.54 min, MH⁺ = 497,Yield: 25 mg, 19%

Example 162: (2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((4-methylmorpholin- 2-yl)methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoate 9prepared from:Inter- mediate 164 and Inter- mediate 11) System B, 0.87 min, MH⁺ = 540,Yield: 84 mg, 59%

Example 163: (2S,3R)- (S)-tetrahydrofuran-3- yl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1-((4- methylmorpholin-2-yl)methyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate(prepared from: Intermediate 164 and Intermediate 40) System B, 0.69min, MH⁺ = 554, Yield: 85 mg, 58%

Example 164: (2S)- cyclopentyl 2-(((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((4-methyl- morpholin-2-yl)methyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)-3-methoxy- propanoate(prepared from: Intermediate 164 and Intermediate 196) System B, 0.92min, MH⁺ = 552, Yield: 13 mg, 9%

Example 165: (2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((4-methylmorpholin- 3-yl)methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Intermediate 167 and Intermediate 11) System B, 0.84 min, MH⁺ = 540,Yield: 30 mg, 21%

Example 166: (2S)- cyclopentyl 2-(((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin- 3-yl)-1-((4-methyl- morpholin-3-yl) methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-3- methoxypropanoate (prepared from:Inter- mediate 167 and Inter- mediate 196) System B, 0.89 min, MH⁺ =552, Yield: 39 mg, 27%

Example 167: (2S,3R)- (S)-tetrahydrofuran-3- yl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1-((4- methylmorpholin-3-yl)methyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate(prepared from: Intermediate 167 and Intermediate 40) System B, 0.66min, MH⁺ = 554, Yield: 11 mg, 8%

Example 168: (2S,3R)- isopropyl 2-(((1-(cyclo- propyl methyl)-2-(1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1H-benzo[d]imidazol-6-yl)methyl)amino)-3- hydroxybutanoate (prepared from: Inter- mediate173 and Inter- mediate 31) System J,

0.94 min, MH⁺ = 467, Yield: 76 mg, 51% Example 169: (2S,3R)- isobutyl2-(((1-(cyclo- propylmethyl)-2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1H-benzo[d]imidazol- 6-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Intermediate 173 and Intermediate 11)

System J, 1.02 min, MH⁺ = 481, Yield: 90 mg, 60% Example 170: (2S,3R)-tert-butyl 2-(((1-(cyclo- propylmethyl)-2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1H-benzo[d]imidazol- 6-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Inter- mediate 173 and(2S,3R)-tert-butyl 2-

amino-3-hydroxy- butanoate hydro- chloride (commer- cially available))System J, 1.01 min, MH⁺ = 481, Yield: 90 mg, 60% Example 171a:(2S,3R)-isopropyl 2- (((2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1- ((tetrahydro-2H- pyran-3-yl)methyl)-1H-benzo[d]imidazol- 6-yl)methyl)amino)- 3-hydroxybutanoate (singlediastereomer of unknown configu- ration at tetrahydro-

pyran centre, Isomer 1) (prepared from: Intermediate 177a andIntermediate 31) System I, 0.53 mins, MH⁺ = 511, Yield: 68 mg, 49%Example 171b: (2S,3R)-isopropyl 2- (((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H- pyran-3-yl)methyl)-1H-benzo[d]imidazol- 6-yl)methyl)amino)- 3-hydroxybutanoate (singlediastereomer of unknown configu- ration at tetrahydro-

pyran centre, Isomer 2) (prepared from: Intermediate 177b andIntermediate 31) System I, 0.51 mins, MH⁺ = 511, Yield: 70 mg, 49%Example 172a: (2S,3R)-cyclobutyl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H- pyran-3-yl)methyl)-1H-benzo[d]imidazol- 6-yl)methyl)amino)- 3-hydroxybutanoate (singlediastereomer of unknown configu- ration at tetrahydro-

pyran centre, Isomer 1) (prepared from: Intermediate 177a andIntermediate 32) System I, 0.55 min, MH⁺ = 523, Yield: 70 mg, 49%Example 172b: (2S,3R)-cyclobutyl 2- (((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H- pyran-3-yl)methyl)-1H-benzo[d]imidazol- 6-yl)methyl)amino)- 3-hydroxybutanoate (singlediastereomer of unknown configu- ration at tetrahydro-

pyran centre, Isomer 2) (prepared from: Intermediate 177b andIntermediate 32) System I, 0.55 min, MH⁺ = 523, Yield: 25 mg, 18%Example 173: (2S,3R)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(tetrahydro-2H- pyran-4-yl)-1H-benzo[d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 131 and Inter- mediate 31) System I,

0.51 min, MH⁺ = 497, Yield: 60 mg, 41% Example 174: (2S,3R)- isopropyl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro-2H- pyran-2-yl)methyl)- 1H-benzo[d]imidazol-6-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate132 and Inter- mediate 31) System J, 0.98 min, MH⁺ = 511,

Yield: 238 mg, 54% Example 175: (2S,3R)- isobutyl 2-(((2-(1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-((tetrahydro-2H-pyran-2-yl)methyl)- 1H-benzo[d]imidazol- 6-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Inter- mediate 132 and Inter- mediate11) System J, 1.05 min, MH⁺ = 525,

Yield: 234 mg, 77% Example 176: (2S,3R)- isobutyl 2-(((2-(1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-(tetrahydro-2H-pyran-4-yl)-1H-benzo [d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoate(prepared from: Inter- mediate 131 and Inter- mediate 11) System I,

0.57 min, MH⁺ = 511, Yield: 55 mg, 72% Example 177: (2S,3R)- tert-butyl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-(tetrahydro-2H-pyran-4-yl)-1H-benzo [d]imidazol-6-yl) methyl)amino)-3- hydroxybutanoate(prepared from: Inter- mediate 131 and (2S,3R)-tert-butyl 2-

amino-3-hydroxy- butanoate hydro- chloride (commer- cially available))System J, 0.90, MH⁺ = 511, Yield: 48 mg, 63% Example 178: (2S,3R)-tert-butyl 2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro-2H- pyran-2-yl)methyl)- 1H-benzo[d]imidazol-6-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate132 and (2S,3R)-tert-butyl 2- amino-3-hydroxy-

butanoate hydro- chloride (commer- cially available)) System J, 1.05min, MH⁺ = 525, Yield: 116 mg, 77% Example 179: (2S,3R)- isopropyl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-(3-methoxy-2-(methoxymethyl) propyl)-1H-benzo[d] imidazol-6-yl)methyl)amino)-3-hydroxy- butanoate (prepared from: Intermediate 134 andIntermediate 31a) System C, 0.53 min, MH⁺ = 515,

Yield: 54 mg, 43% Example 180: (2S,3R)- (S)-tetrahydrofuran-3- yl2-(((2-(1,5-dimethyl- 6-oxo-1,6-dihydro- pyridin-3-yl)-1-((tetra-hydro-2H-pyran-2-yl) methyl)-1H-benzo [d]imidazol-6-yl)methyl)amino)-3-hydroxy- butanoate (prepared from: Intermediate 132 andIntermediate 40) System I, 0.51 min,

MH⁺ = 539, Yield: 147 mg, 63% Example 181: (2S,3R)- isopropyl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-(1-(tetrahydro-2H- pyran-4-yl)ethyl)-1H- benzo[d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoate (prepared from: Inter- mediate 133 andInter- mediate 31) System I, 0.55 min, MH⁺ = 525, Yield: 111 mg, 25%

Example 182: (2S,3R)- tert-butyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1- (1-(tetrahydro-2H- pyran-4-yl)ethyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 133 and (2S,3R)-tert-butyl 2- amino-3-hydroxy- butanoatehydro- chloride (commer-

cially available)) System I, 0.59 min, MH⁺ = 539, Yield: 65 mg, 62%Example 183: (2S,3R)- tert-butyl 2-(((1-(((R)- 1-acetylpiperidin-3-yl)methyl)-2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxybutanoate (preparedfrom: Inter- mediate 136 and (2S,3R)-tert-butyl 2- amino-3-hydroxy-butanoate hydro- chloride (commer- cially available)) System I, 0.55min, MH⁺ = 566, Yield

53 mg, 36% Example 184: (S)- cyclobutyl 4-methyl- 2-(((2-(5-methyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) pentanoate (prepared from:Intermediate 151 and Intermediate 20) System B, 1.08 min, MH⁺ = 521,Yield: 31 mg, 28%

Example 185: (S)- tetrahydro-2H-pyran- 4-yl 4-methyl-2-(((2-(5-methyl-6-oxo-l,6- dihydropyridin-3-yl)- 1-((tetrahydro-2H-pyran-4-yl)methyl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino) pentanoate(prepared from: Intermediate 151 and Intermediate 16) System B, 0.96min, MH⁺ = 551, Yield: 61 mg, 52%

Example 186: (S)- neopentyl 3-methyl- 2-(((2-(5-methyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoate (prepared from:Intermediate 151 and Intermediate 15) System B, 1.16 min, MH⁺ = 523,

Yield: 56 mg, 50% Example 187: (2S)- 1-methoxypropan-2- yl4-methyl-2-(((2- (5-methyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino) pentanoate (prepared from: Intermediate 151 andIntermediate 18) System B, 1.0 min, MH⁺ = 539, Yield: 46 mg, 40%

Example 189: (2S,3R)- neopentyl 3-hydroxy- 2-(((2-(5-methyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoate (prepared from:Intermediate 151 and Intermediate 10) System B, 0.93 min, MH⁺ = 525,Yield: 35 mg, 31%

Example 190: (S)- isopropyl 4-methyl- 2-(((2-(5-methyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) pentanoate (prepared from:Intermediate 151 and Intermediate 21) System B, 1.07 min, MH⁺ = 509,Yield: 60 mg, 55%

Example 191: (2S,3R)- neopentyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxybutanoate (preparedfrom: Inter- mediate 116 and Inter- mediate 10) System B, 0.97 min, MH⁺= 539, Yield: 82 mg, 74%

Example 192: (S)- cyclopentyl 3-(1H- imidazol-5-yl)-2-(((2-(5-methyl-6-oxo- 1,6-dihydropyridin-3- yl)-1-((tetrahydro-2H-pyran-4-yl) methyl)-1H-benzo[d] imidazol-5-yl)methyl)amino)propanoate (prepared from: Inter- mediate 116 and Inter- mediate286) System B, 0.79 min, MH⁺ = 558, Yield: 57 mg, 24%

Example 193: (S)-neo- pentyl 3-methoxy-2- (((2-(5-methyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) propanoate (prepared from:Intermediate 151 and Intermediate 192) System B, 0.96 min, MH⁺ = 525,Yield: 70 mg, 63%

Example 194: (S)- cyclopentyl 3- methoxy-2-(((2-(5- methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(oxetan-3- ylmethyl)-1H-benzo [d]imidazol-5-yl)methyl)amino) propanoate (prepared from: Intermediate 195 andIntermediate 196) System B, 0.85 min, MH⁺ = 495, Yield: 69 mg, 61%

Example 195: (S)- cyclopentyl 2-(((2- (1,5-dimethyl-6- oxo-1,6-dihydro-pyridin-3-yl)-1- ethyl-1H-benzo[d] imidazol-5-yl) methyl)amino)-3-methoxypropanoate (prepared from: Inter- mediate 198 and Inter- mediate196) System B, 0.99 min, MH⁺ =

467, Yield: 155 mg, 73% Example 196: (2S,3R)- cyclopentyl 2-(((2-(1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-ethyl-1H-benzo[d]imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate (prepared from:Intermediate 198 and Intermediate 137) System B, 0.94 min, MH⁺ = 467,Yield: 167 mg, 78%

Example 197: (2S,3R)- cyclopentyl 3-hydroxy- 2-(((1-(2-methoxyethyl)-2-(5-methyl-6-oxo-1,6- dihydropyridin-3-yl)- 1H-benzo[d]imidazol-5-yl)methyl)amino) butanoate (prepared from: Intermediate 201 andIntermediate 137) System B, 0.87 min, MH⁺ = 483, Yield: 141 mg, 67%

Example 198: (S)-(S)- tetrahydrofuran-3-yl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1-ethyl- 1H-benzo[d]imidazol-5-yl)methyl)amino)- 4-methylpentanoate (prepared from: Inter- mediate198 and Inter- mediate 30) System B, 0.99 min, MH⁺ = 481, Yield: 154 mg,70%

Example 199: (2S,3R)- neopentyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-ethyl-1H-benzo[d] imidazol-5-yl)methyl)amino)-3-hydroxy- butanoate (prepared from: Intermediate 198 andIntermediate 10) System B, 1.01 min, MH⁺ = 469, Yield: 153 mg, 71%

Example 200: (2S,3R)- neopentyl 3-hydroxy- 2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin- 3-yl)-1-(oxetan-3- ylmethyl)-1H-benzo[d]imidazol-5-yl)methyl) amino)butanoate (prepared from: nd Intermediate195 and Intermediate 10) System B, 0.87 min, MH⁺ = 497, Yield: 87 mg,77%

Example 201: (S)- cyclopentyl 2-(((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin- 3-yl)-1-(2-methoxy- ethyl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-3- methoxypropanoate (prepared from: Inter-mediate 203 and Inter- mediate 196) System B, 0.97 min, MH⁺ = 497,Yield: 125 mg, 61%

Example 202: (2S,3R)- cyclopentyl 2-(((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin- 3-yl)-1-(2-methoxy- ethyl)-1H-benzo[d]imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate (prepared from:Intermediate 203 and Intermediate 137) System B, 0.91 min, MH⁺ = 497,Yield: 156 mg, 76%

Example 203: (2S,3R)- cyclopentyl 2-(((1-(2- (dimethylamino)ethyl)-2-(5-methyl-6-oxo-1,6- dihydropyridin-3-yl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoate (preared from: Inter- mediate 206and Inter- mediate 137) System B, 0.86 min, MH⁺ = 496, Yield: 163 mg,79%

Example 204: (2S,3R)- neopentyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(2-methoxyethyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate203 and Inter- mediate 10) System B, 0.98 min, MH⁺ = 499, Yield: 133 mg,64%

Example 205: (2S,3R)- neopentyl 2-(((1-(2- (dimethylamino)ethyl)-2-(5-methyl-6-oxo-1,6- dihydropyridin-3-yl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate206 and Inter- mediate 10) System B, 0.92 min, MH⁺ = 498, Yield: 164 mg,79%

Example 206: (2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-ethyl-1H-benzo[d] imidazol-5-yl)methyl)amino)-3-hydroxy- butanoate (prepared from: Intermediate 198 andIntermediate 11) System B, 0.93 min, MH⁺ = 455, Yield: 59 mg, 51%

Example 207: (S)- cyclopentyl 3- methoxy-2-(((1- methyl-2-(5-methyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1H- benzo[d]imidazol-5-yl)methyl)amino) propanoate (pre- pared from: Inter- mediate 208 andIntermediate 196)

System B, 0.89 min, MH⁺ = 439, Yield: 18 mg, 7.3% Example 208: (2S,3R)-neopentyl 3-hydroxy- 2-(((1-methyl-2-(5- methyl-6-oxo-1,6-dihydropyridin-3-yl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoate(prepared from: Intermediate 208 and Intermediate 10) System B, 0.90min, MH⁺ = 441, Yield: 24 mg, 9.7%

Example 209: (2S,3R)- isobutyl 3-hydroxy-2- (((1-methyl-2-(5-methyl-6-oxo-1,6- dihydropyridin-3-yl)- 1H-benzo[d]imidazol-5-yl)methyl)amino) butanoate (prepared from: Intermediate 208 andIntermediate 11) System B, 0.84 min, MH⁺ = 427, Yield: 24 mg, 10%

Example 210: (S)-(S)- tetrahydrofuran-3-yl 2- (((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1-ethyl- 1H-benzo[d]imidazol-5-yl)methyl)amino)- 3-methylbutanoate (prepared from: Inter- mediate 198and Inter- mediate 283) System

B, 0.93 min, MH⁺ = 467, Yield: 15 mg, 7% Example 211: (2S,3R)- isobutyl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-(2-methoxyethyl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Inter- mediate 203 and Inter- mediate11) System B, 0.91 min, MH⁺ = 485, Yield: 106 mg, 53%

Example 212: (2S,3R)- tert-butyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxybutanoate (preparedfrom: Inter- mediate 116 and (2S,3R)-tert-butyl 2- amino-3-hydroxy-butanoate hydro- chloride (commer- cially available)) System B, 0.89min,

MH⁺ = 525, Yield: 67 mg, 62% Example 213: (2S,3R)- isobutyl 2-(((2-(1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-((tetrahydro-2H-pyran-3-yl)methyl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Inter- mediate 212 and Inter- mediate11) System C, 0.59, MH⁺ = 525, Yield: 88 mg, 45%

Example 214: (2S,3R)- (S)-tetrahydrofuran-3- yl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo [d]imidazol-5-yl) methyl)amino)-3-hydroxybutanoate (prepared from: Inter- mediate 116 and Inter- mediate40) System B, 0.71 min, MH⁺ = 539, Yield: 3.2 mg, 4.3%

Example 215: (2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((S)-1-methoxy- propan-2-yl)-1H-benzo[d]imidazol-5- yl)methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 215 and Inter- mediate 11) System B, 0.96 min, MH⁺ = 499,Yield: 53 mg, 48%

Example 216: (2S,3R)- (S)-tetrahydrofuran-3- yl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1-((S)-1- methoxypropan-2-yl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxybutanoate ¹H NMR(d₆-DMSO): δ 1.12 (d, J = 6.3 Hz, 3 H), 1.53 (d, J = 7.1 Hz, 3 H),1.79-1.87 (m, 1 H), 2.07-2.16 (m, 4 H), 3.15 (s, 3 H), 3.54 (s, 3 H),3.65- 3.87 (m, 9 H), 3.91 (d, J = 13.1 Hz, 1 H), 4.00 (t, J = 9.9 Hz, 1H), 4.69-4.80 (m,

2 H), 5.21-5.27 (m, 1 H), 7.18 (dd, J = 8.3, 1.3 Hz, 1 H), 7.54 (s, 1H), 7.60-7.65 (m, 1 H), 7.69 (d, J = 8.3 Hz, 1 H), 8.00 (d, J = 2.3 Hz,1 H). (pre- pared from: Interme- diate 215 and Inter- mediate 40) SystemB, 0.76 min, no MH+ Example 217: (S)- cyclopentyl 2-(((2-(1,5-dimethyl-6- oxo-1,6-dihydro- pyridin-3-yl)-1-((S)- 1-methoxypropan-2-yl)-1H-benzo[d] imidazol-5-yl) methyl)amino)-3- methoxypropanoate(prepared from: Inter- mediate 215 and Inter- mediate 196) System B,1.02 min, MH⁺ = 511, Yield: 152 mg, 72%

Example 218: (2S,3R)- cyclopentyl 2-(((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((S)-1-methoxy- propan-2-yl)-1H-benzo[d]imidazol-5- yl)methyl)amino)- 3-hydroxybutanoate (prepared from:Inter- mediate 215 and Inter- mediate 137) System B, 0.96, MH⁺ = 511,Yield: 121 mg, 57%

Example 219: (2S,3R)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((S)-1-methoxy- propan-2-yl)-1H-benzo[d]imidazol-5- yl)methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 215 and Inter- mediate 31) System B, 0.88 min, MH⁺ = 485,Yield: 56 mg, 52%

Example 220: (2S,3R)- (S)-sec-butyl 2-(((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxybutanoate (preparedfrom: Inter- mediate 116 and Inter- mediate 34) System B, 0.89 min, MH⁺= 525, Yield: 34 mg, 48%

Example 221: (2S,3R)- cyclobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxybutanoate (preparedfrom: Inter- mediate 116 and Inter- mediate 32) System B, 0.90 min, MH⁺= 523, Yield: 232 mg, 65%

Example 222: (2S,3R)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((R)-1-methoxy- propan-2-yl)-1H-benzo[d]imidazol-5-yl) methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 218 and Inter- mediate 31) System B, 0.87 min, MH⁺ = 485,Yield: 61 mg, 61%

Example 223: (2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((R)-1-methoxy- propan-2-yl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxybutanoate (prepared from:Inter- mediate 218 and Inter- mediate 11) System B, 0.96 min, MH⁺ = 499,Yield: 80 mg, 52%

Example 224: (2S,3R)- (S)-tetrahydrofuran-3- yl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1-(((S)- tetrahydrofuran-2-yl)methyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)-3-hydroxy- butanoate(prepared from: Intermediate 114 and Intermediate 40) System B, 0.75min, MH⁺ = 525, Yield: 231 mg, 77%

Example 225: (2R,3S)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-3-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxybutanoate (preparedfrom: Inter- mediate 212 and Inter- mediate 31) System J, 0.91 min, MH⁺= 511, Yield: 2.08 g 72%

Example 226: (2S,3R)- cyclobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(((S)-tetrahydro- furan-2-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxybutanoate (preparedfrom: Inter- mediate 114 and Inter- mediate 32) System J, 0.90 min, MH⁺= 509, Yield: 50 mg, 14%

Example 227: (2S,3R)- tert-butyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((S)-1-methoxy- propan-2-yl)-1H-benzo[d]imidazol-5- yl)methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 215 and (2S,3R)-tert-butyl 2- amino-3-hydroxy- butanoatehydro- chloride (commer-

cially available)) System J, 0.99 min, MH⁺ = 499, Yield: 21 mg, 24%Example 228: (2S,3R)- cyclobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydrofuran- 3-yl)methyl)-1H-benzo[d]imidazol-5- yl)methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 221 and Inter- mediate 32) System J, 0.88 min, MH⁺ = 509,Yield: 312 mg, 60%

Example 229: (2S,3R)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydrofuran-3- yl)methyl)-1H-benzo[d]imidazol-5-yl) methyl)arnino)-3- hydroxybutanoate (prepared from:Inter- mediate 221 and Inter- mediate 31) System J, 0.85 min, MH⁺ = 497,Yield: 236 mg, 42%

Example 230: (2S,3R)- cyclobutyl 2-(((1-(1,3- dimethoxypropan-2-yl)-2-(1,5-dimethyl- 6-oxo-1,6-dihydro- pyridin-3-yl)-1H-benzo[d]imidazol-5- yl)methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 224 and Inter- mediate 32) System J, 0.95 min, MH⁺ = 529,

Yield: 147 mg, 76% Example 231: (2S,3R)- tert-butyl 2-(((1-(1,3-dimethoxypropan-2- yl)-2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1H-benzo [d]imidazol-5-yl) methyl)amino)-3-hydroxybutanoate (prepared from: Inter- mediate 224 and(2S,3R)-tert-butyl 2- amino-3-hydroxy-

butanoate hydro- chloride (commer- cially available)) System J, 0.98min, MH⁺ = 529, Yield: 67 mg, 47% Example 232: (2S,3R)- tert-butyl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydrofuran-3- yl)methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoate (prepared from: Inter- mediate 221 and(2S,3R)-tert-butyl 2- amino-3-hydroxy- butanoate hydro- chloride(commer-

cially available)) System J, 0.91 min, MH⁺ = 511, Yield: 93 mg, 40%Example 233: (S)- cyclopentyl 2-(((2- (1,5-dimethyl-6- oxo-1,6-dihydro-pyridin-3-yl)-1- ((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxypropanoate 9preparingfrom: Intermediate 116 and Intermediate 276) System J, 0.89 min, MH⁺ =523,

Yield: 142 mg, 76% Example 234: (2S)- tetrahydrofuran-3- yl2-(((2-(5-methyl- 6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H-pyran-4-yl)methyl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoate(prepared from: Intermediate 151 and Intermediate 35) System B, 0.90min, MH⁺ = 509,

Yield: 154 mg, 89% Example 235: (2S)- 1-methoxypropan-2- yl2-(((2-(5-methyl- 6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H-pyran-4-yl)methyl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoate(prepared from: Intermediate 151 and Intermediate 37) System B, 0.86min, MH⁺ = 511,

Yield: 140 mg, 80% Example 236: (2S)- tetrahydrofuran-3-yl3-methyl-2-(((2-(5- methyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro- 2H-pyran-4-yl) methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino) butanoate (prepared from: Intermediate 151 andIntermediate 5) System B, 0.88

min, MH⁺ = 523, Yield: 84 mg, 47% Example 237: (2S)- 1-methoxypropan-2-yl 3-methyl-2-(((2- (5-methyl-6-oxo- 1,6-dihydropyridin-3-yl)-1-((tetrahydro- 2H-pyran-4-yl) methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino) butanoate (prepared from: Intermediate 151 andIntermediate 41) System B, 0.96

min, MH⁺ = 525, Yield: 96 mg, 54% Example 238: (2S)- 1-methoxypropan-2-yl 3,3-dimethyl- 2-(((2-(5-methyl- 6-oxo-1,6-dihydro- pyridin-3-yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)- 1H-benzo[d] imidazol-5-yl)methyl)amino) butanoate (prepared from: Intermediate 151 andIntermediate 43) System B, 1.05

min, MH⁺ = 539, Yield: 92 mg, 50% Example 239: (2S)-tetrahydrofuran-3-yl 3,3-dimethyl-2-(((2- (5-methyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((tetrahydro- 2H-pyran-4-yl) methyl)-1H-benzo[d]imidazol-5-yl) methyl)amino) butanoate (prepared from Intermediate151 and Intermediate 45) System B, 0.96 min, MH⁺ = 537,

Yield: 130 mg, 71% Example 240: (2S)- tetrahydrofuran-3- yl 3-hydroxy-3-methyl-2-(((2-(5- methyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino) butanoate (prepared from: Intermediate 151 andIntermediate 47) System B, 0.73 min, MH⁺ = 539,

Yield: 33 mg, 18% Example 241: (S)- (S)-tetrahydrofuran- 3-yl3-methyl-2- (((2-(5-methyl-6- oxo-1,6-dihydro- pyridin-3-yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino) butanoate (prepared from: Intermediate 151) andIntermediate 49) System B, 0.87

min, MH⁺ = 523, Yield: 99 mg, 56% Example 242: (S)- ethyl 3-cyclopropyl-2-(((2-(5-methyl- 6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H-pyran-4-yl)methyl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino) propanoate(prepared from: Intermediate 151 and (S)-ethyl 2-amino-3-cyclo-propylpropanoate, Hydrochloride (com- mercially available))

System B, 0.93 min, MH⁺ = 493, Yield: 37 mg, 53% Example 243: (S)-cyclopentyl 3- hydroxy-3-methyl- 2-(((2-(5-methyl-6- oxo-1,6-dihydro-pyridin-3-yl)-1- ((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoate (prepared from:Intermediate 151 and Intermediate 47) System B, 0.93 min, MH⁺ = 537,Yield: 36 mg, 20%

Example 244: (2S)- 1-methoxypropan-2- yl 3-hydroxy-3- methyl-2-(((2-(5-methyl-6-oxo-1,6- dihydropyridin-3- yl)-1-((tetrahydro- 2H-pyran-4-yl)methyl)-1H-benzo [d]imidazol-5-yl) methyl)amino) butanoate (pre- paredfrom: Inter- mediate 151 and Intermediate 53) System B, 0.81

min, MH⁺ = 541, Yield: 42 mg, 23% Example 245: (S)- cyclopentyl 2-cyclopropyl-2-(((2- (5-methyl-6-oxo- 1,6-dihydro- pyridin-3-yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino) acetate (Prepared from: Intermediate 151 andIntermediate 55) System B, 1.00 min, MH⁺ = 519,

Yield: 61 mg, 60% Example 246: (S)- cyclopentyl 3-cyclo-propyl-2-(((2-(5- methyl-6-oxo-1,6- dihydropyridin-3-yl)-1-((tetrahydro- 2H-pyran-4-yl) methyl)-1H-benzo [d]imidazol-5-yl)methyl)amino) propanoate (pre- pared from: Inter- mediate 151 andIntermediate 57) System B, 1.08 min, MH⁺ = 533, Yield:

48 mg, 45% Example 247: (R)- cyclopentyl 3-cyclo- propyl-2-(((2-(5-methyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-((tetrahydro-2H-pyran-4-yl)methyl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino) propanoate(prepared from: Intermediate 151 and Intermediate 59) System B, 1.08min, MH⁺ = 533, Yield: 57 mg, 54%

Example 248: (2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-2-yl)methyl)-1H-benzo[d]imidazol- 5-yl)methyl)amino)- 3-hydroxybutanoate (preparedfrom: Inter- mediate 225 and Inter- mediate 11) System A, 0.67 min, MH⁺= 525, Yield: 138 mg, 58%

Example 249: (2S,3R)- isobutyl 2-((2-((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-ethyl-1H-benzo[d] imidazol-5-yl)oxy)ethyl)amino)-3- hydroxybutanoate (prepared from: Inter- mediate 228 andInter- mediate 11) System B,

0.96 min, MH⁺ = 485, Yield: 4.3 mg, 5.9% Example 250: (2S,3R)- isobutyl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-((R)-1-hydroxy-propan-2-yl)-1H- benzo[d]imidazol-5- yl)methyl)amino)-3-hydroxybutanoate (prepared from: Inter- mediate 233 and Inter- mediate11) System B, 0.85 min, MH⁺ = 485,

Yield: 59 mg, 40% Example 251: (2S,3R)- isopropyl 2-(((2-(1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-((R)-1-hydroxy-propan-2-yl)-1H-benzo [d]imidazol-5-yl) methyl)amino)-3-hydroxybutanoate (prepared from: Inter- mediate 233 and Inter- mediate31) System B, 0.77 min, MH⁺ = 471, Yield: 80 mg, 55%

Example 252: (S)- cyclopentyl 2-((2-((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin- 3-yl)-1-ethyl-1H- benzo[d]imidazol- 5-yl)oxy)ethyl)amino)-4-methoxy- butanoate (prepared from: Intermediate 228 andIntermediate 23) System B, 1.04 min, MH⁺ = 511, Yield: 12 mg, 10%

Example 253: (S)- cyclopentyl 2-((2-((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin- 3-yl)-1-((tetrahydro- 2H-pyran-4-yl)methyl)-1H-benzo [d]imidazol-5-yl) oxy)ethyl)amino)-3- methoxypropanoate(prepared from: Inter- mediate 261 and Inter- mediate 196) System B,0.99 min, MH⁺ = 567, Yield: 9 mg, 16%

Example 254: (S)- cyclopentyl 2-((2- ((2-(1,5-dimethyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1- ((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d] imidazol-5-yl)oxy) ethyl)amino) propanoate (pre- pared from:Inter- mediate 261 and Inter-

mediate 61) System B, 1.00 min, MH⁺ = 537, Yield: 15 mg, 13% Example255: (2S,3R)- isopropyl 2-((2-((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin-3- yl)-1-ethyl-1H- benzo[d]imidazol-5-yl)oxy)ethyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate228 and Inter- mediate 31) System B,

0.88 min, MH⁺ = 471, Yield: 12 mg, 10% Example 256: (S)-(S)-tetrahydrofuran-3-yl 2-((2-((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-ethyl-1H-benzo[d] imidazol-5-yl)oxy)ethyl)amino)-3- methylbutanoate (prepared from: Inter- mediate 228 andInter-

mediate 49) System B, 0.95 min, MH⁺ = 497, Yield: 13 mg, 11% Example257: (2S,3R)- isobutyl 2-((2-((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)oxy)ethyl) amino)-3-hydroxy- butanoate(prepared from: Intermediate 261 and Intermediate 11) System B, 0.93min, MH⁺ = 555, Yield: 21 mg, 18%

Example 258: (2S,3R)- isopropyl 2-((2-((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin- 3-yl)-1-((tetrahydro- 2H-pyran-4-yl)methyl)-1H-benzo[d] imidazol-5-yl)oxy) ethyl)amino)-3- hydroxybutanoate(prepared from: Inter- mediate 261 and Inter- mediate 31) System B, 0.85min, MH⁺ = 541, Yield: 24 mg,

21% Example 259: (2S,3R)- isobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-((S)-1-hydroxy- propan-2-yl)-1H-benzo[d]imidazol-5- yl)methyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 236 and Inter- mediate 11) System A, 0.51 min, MH⁺ =

485, Yield: 50 mg, 34% Example 260: (S)-(S)- tetrahydrofuran-3-yl2-(((2-(1,5- dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-((R)-1-hydroxy-propan-2-yl)-1H- benzo[d]imidazol-5- yl)methyl)amino)-3- methylbutanoate(prepared from: Inter- mediate 233 and Inter- mediate 49) System

B, 0.83 min, MH⁺ = 497, Yield: 47 mg, 31% Example 261: (S)-(S)-tetrahydrofuran-3-yl 2-(((2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1-(2- hydroxyethyl)-1H benzo[d]imidazol-5-yl)methyl)amino)-3- methylbutanoate (prepared from: Inter- mediate 278and Inter- mediate 49) System 1, 0.41 min, MH⁺ = 483,

Yield: 50 mg, 32% Example 262: (2S,3R)- isobutyl 3-hydroxy-2-(((2-(5-methyl-6-oxo- 1,6-dihydropyridin-3- yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino) butanoate(prepared from: Intermediate 245 and Intermediate 11) System 1, 0.62min, MH⁺ = 511, Yield: 49 mg, 34%

Example 263: (S)- isopropyl 2-(((2- (1,5-dimethyl-6- oxo-1,6-dihydro-pyridin-3-yl)-1-((S)- 1-hydroxypropan- 2-yl)-1H-benzo[d] imidazol-5-yl)methyl)amino)-3- methylbutanoate (prepared from: Inter- mediate 236 andInter- mediate 12) System I,

0.54 min, MH⁺ = 469, Yield: 58 mg, 40% Example 264: (S)-(S)-tetrahydrofuran-3-yl 2-(((2-(1,5-dimethyl- 6-oxo-1,6-dihydro-pyridin-3-yl)-1-((S)- 1-hydroxypropan-2- yl)-1H-benzo[d] imidazol-5-yl)methyl)amino)-3- methylbutanoate (prepared from: Inter- mediate 236 andInter- mediate 49) System 1,

0.46 min, MH⁺ = 497, Yield: 26 mg, 17% Example 265a: tetra-hydrofuran-3-yl 2- (((2-(1,5-dimethyl-6- oxo-1,6-dihydro-pyridin-3-yl)-1-((S)- 1-hydroxypropan-2- yl)-1H-benzo[d]imidazol-5-yl)methyl) amino)-3-methoxy- butanoate Diastereo- isomer 1(prepared from: Intermediate 236 and Intermediate 9) System I, 0.44 min,MH⁺ = 513,

Yield: 4.4 mg, 2.8% Example 265b: tetra- hydrofuran-3-yl 2-(((2-(1,5-dimethyl-6- oxo-1,6-dihydro- pyridin-3-yl)-1-((S)-1-hydroxypropan-2- yl)-1H-benzo[d] imidazol-5-yl)methyl)amino)-3-methoxy- butanoate Diastereo- isomer 2 (prepared from:Intermediate 236 and Intermediate 9) System J, 0.77 min, MH⁺ = 513,

Yield: 35 mg, 22% Example 266: (2S,3R)- isopropyl 2-(((2-(1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-((tetrahydro-2H-pyran-2-yl)methyl)- 1H-benzo[d]imidazol- 5-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Inter- mediate 225 and Inter- mediate31) System 1, 0.60 min, MH⁺ = 511, Yield: 85 mg, 30%

Example 267: (2S,3R)- isopropyl 3-hydroxy- 2-(((2-(5-methyl-6-oxo-1,6-dihydro- pyridin-3-yl)-1-((tetra- hydro-2H-pyran-2-yl)methyl)-1H-benzo[d] imidazol-5-yl)methyl) amino)butanoate (preparedfrom: Inter- mediate 245 and Inter- mediate 31 System 1, 0.58 min, MH⁺ =497, Yield: 132 mg, 47%

Example 268: (2S,3R)- isopropyl 2-((2-((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((S)-1-methoxy- propan-2-yl)-1H-benzo[d]imidazol-5-yl)oxy) ethyl)amino)-3- hydroxybutanoate (prepared from:Inter- mediate 247 and Inter- mediate 31) System J, 0.97 min, MH⁺ = 515,Yield: 7 mg, 7%

Example 269: (S)- isobutyl 2-((2-((2- (1,5-dimethyl-6- oxo-1,6-dihydro-pyridin-3-yl)-1-((S)- 1-methoxypropan-2- yl)-1H-benzo[d]imidazol-5-yl)oxy) ethyl)amino)-3- methoxypropanoate (prepared from:Inter- mediate 247 and Inter- mediate 66) System J, 1.09 min, MH⁺ = 529,Yield: 8 mg, 8%

Example 270: (2S,3R)- isopropyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(2-methoxypropyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate256 and Inter- mediate 31) System I, 0.54 min, MH⁺ = 485, Yield: 36 mg,13%

Example 271: (2S,3R)- neopentyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(2-methoxypropyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate256 and Inter- mediate 10) System I, 0.65 min, MH⁺ = 513, Yield: 311 mg,82%

Example 272: (2S,3R)- tert-butyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(2-methoxypropyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate256 and (2S,3R)-tert-butyl 2-amino-3-hydroxy- butanoate hydro- chloride(commer-

cially available)) System 1, 0.59 min, MH⁺ = 499, Yield: 311 mg, 82%Example 273: (2S,3R)- cyclobutyl 2-(((2-(1,5- dimethyl-6-oxo-1,6-dihydropyridin-3-yl)- 1-(2-methoxypropyl)- 1H-benzo[d]imidazol-5-yl)methyl)amino)- 3-hydroxybutanoate (prepared from: Inter- mediate256 and Inter- mediate 32) System I, 0.57 min, MH⁺ = 497, Yield: 290 mg,66%

Example 274: (2S,3R)- tert-butyl 2-((2-((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)oxy)ethyl) amino)-3-hydroxy butanoate(prepared from: Intermediate 261 and (2S,3R)-tert- butyl 2-amino-3-hydroxybutanoate hydrochloride (com-

mercially available)) System J, 0.96 min, MH⁺ = 545, Yield: 25 mg, 17%Example 275: (2S,3R)- cyclobutyl 2-((2-((2- (1,5-dimethyl-6-oxo-1,6-dihydropyridin-3- yl)-1-((tetrahydro-2H- pyran-4-yl)methyl)-1H-benzo[d]imidazol- 5-yl)oxy)ethyl)amino)- 3-hydroxy butanoate(prepared from: Inter- mediate 261 and Inter- mediate 32) System J, 0.93min, MH⁺ = 553, Yield: 9.2 mg, 3.3%

Example 276: (2S,3R)- (S)-tetrahydrofuran-3- yl 2-((2-((2-(1,5-dimethyl-6-oxo-1,6- dihydropyridin-3-yl)- 1-((tetrahydro-2H-pyran-4-yl)methyl)- 1H-benzo[d]imidazol- 5-yl)oxy)ethyl)amino)-3-hydroxy butanoate (prepared from: Inter- mediate 261 and Inter-mediate 40) System J, 0.79 min, MH⁺ = 569, Yield: 170 mg, 12%

Example 108a and Example 108b: (2S,3R)-Isopropyl3-hydroxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((-tetrahydro-2-pyran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoate(Isomer 1) and (2S,3R)-isopropyl3-hydroxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((-tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoate(Isomer 2)

The racemic (2S,3R)-Isopropyl3-hydroxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((-tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoate(For a preparation see Example 108, 120 mg) was separated by chiralHPLC. The HPLC purification was carried out on a Chiralcel OD-H column(Lot. No. ODH11158-01, 250×30 mm). The purification was run using 30%EtOH in heptane, with a flow rate of 30 mL/min. UV detection was at 215nm. The first eluting enantiomer was collected, and the fractionsevaporated under reduced pressure, to give the title compound “Isomer1”, Example 108a (41 mg). The second eluting enantiomer was collected,and the fractions evaporated under reduced pressure, to give the titlecompound “Isomer 2”, Example 108b (38 mg). Isomer 1: LCMS (System J):t_(RET)=0.85 min; MH⁺ 497. Isomer 2: LCMS (System J): t_(RET)=0.85 min;MH⁺ 497.

Example 130a: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer1)

Example 130b: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer2)

A sample of (rac)-(2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(for a preparation see Example 130, 290 mg) was separated by chiralchromatography using a 30 mm×25 cm Chiralcel OD-H column, eluting with60% EtOH/Heptane at a flowrate of 30 mL/min. The appropriate fractionsfor each isomer were combined and evaporated under reduced pressure togive the title compounds

Example 130a: Isomer 1 (107 mg)

LCMS (System I): t_(RET)=0.59 min; MH⁺ 499. Chiral analysis carried outon a 4.6 mmid×25 cm Chiralcel OD-H column, eluting with 60% EtOH/Heptaneat a flowrate of 1 mL/min, chiral purity >99.5%.

Example 130b: Isomer 2 (96 mg)

LCMS (System I): t_(RET)=0.59 min; MH⁺ 499. Chiral analysis carried outon a 4.6 mmid×25 cm Chiralcel OD-H column, eluting with 60% EtOH/Heptaneat a flowrate of 1 mL/min, chiral purity=98.5%.

Example 131a: (2S,3R)-cyclobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer1)

Example 131b: (2S,3R)-cyclobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer2)

A sample of (rac)-(2S,3R)-cyclobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(for a preparation see Example 131, 300 mg) was separated by chiralchromatography using a 30 mm×25 cm Chiralcel OD-H column, eluting with10% EtOH/Heptane at a flowrate of 30 mL/min. The appropriate fractionsfor each isomer were combined and evaporated under reduced pressure togive the title compounds

Example 131a: Isomer 1 (58 mg)

LCMS (System I): t_(RET)=0.61 min; MH⁺ 511. Chiral analysis carried outon a 4.6 mmid×25 cm Chiralcel OD-H column, eluting with 10% EtOH/Heptaneat a flowrate of 1 mL/min, chiral purity=97.3%%.

Example 131b: Isomer 2 (92 mg)

LCMS (System I): t_(RET)=0.61 min; MH⁺ 511. Chiral analysis carried outon a 4.6 mmid×25 cm Chiralcel OD-H column, eluting with 10% EtOH/Heptaneat a flowrate of 1 mL/min, chiral purity >99.5%.

Example 132a: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer1)

Example 132b: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer2)

A sample of (rac)-(2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methoxybutan-2-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(for a preparation see Example 132, 50 mg) was separated by chiralchromatography using a 30 mm×25 cm Chiralcel OD-H column, eluting with70% EtOH/Heptane at a flowrate of 30 mL/min. The appropriate fractionsfor each isomer were combined and evaporated under reduced pressure togive the title compounds

Example 132a: Isomer 1 (18 mg)

LCMS (System I): t_(RET)=0.62 min; MH⁺ 513. Chiral analysis carried outon a 4.6 mmid×25 cm Chiralcel OD-H column, eluting with 70% EtOH/Heptaneat a flowrate of 1 mL/min, chiral purity=97.3%.

Example 132b: Isomer 2 (14 mg)

LCMS (System I): t_(RET)=0.62 min; MH⁺ 513. Chiral analysis carried outon a 4.6 mmid×25 cm Chiralcel OD-H column, eluting with 10% EtOH/Heptaneat a flowrate of 1 mL/min, chiral purity >99.5%.

Example 133a: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1-H-benzo[d]imidazole-6-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer1)

Example 133b: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1-H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer2)

A sample of (rac)-(2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate(for a preparation see Example 133, 285 mg) was separated by chiralchromatography using a 30 mm×25 cm Chiralcel OD-H column, eluting with50% EtOH/Heptane at a flowrate of 25 mL/min. The appropriate fractionsfor each isomer were combined and evaporated under reduced pressure togive the title compounds

Example 133a: Isomer 1 (159 mg)

LCMS (System J): t_(RET)=0.84 min; MH⁺ 497. Chiral analysis carried outon a 4.6 mmid×25 cm Chiralcel OD-H column, eluting with 70% EtOH/Heptaneat a flowrate of 1 mL/min, chiral purity >99.5%.

Example 133b: Isomer 2 (145 mg)

LCMS (System J): t_(RET)=0.84 min; MH⁺ 497. Chiral analysis carried outon a 4.6 mmid×25 cm Chiralcel OD-H column, eluting with 70% EtOH/Heptaneat a flowrate of 1 mL/min, chiral purity=99.2%.

Example 134a: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazole-6-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer1)

Example 134b: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer2)

A sample of (rac)-(2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate(for a preparation see Example 134, 200 mg) was separated by chiralchromatography using a 30 mm×25 cm Chiralcel OD-H column, eluting with50% EtOH/Heptane at a flowrate of 25 mL/min. The appropriate fractionsfor each isomer were combined and evaporated under reduced pressure togive the title compounds.

Example 134a: Isomer 1 (100 mg)

LCMS (System J): t_(RET)=0.84 min; MH⁺ 511. Chiral analysis carried outon a 4.6 mmid×25 cm Chiralcel OD-H column, eluting with 70% EtOH/Heptaneat a flowrate of 1 mL/min, chiral purity >99.5%.

Example 134b: Isomer 2 (85 mg)

LCMS (System J): t_(RET)=0.91 min; MH⁺ 511. Chiral analysis carried outon a 4.6 mmid×25 cm Chiralcel OD-H column, eluting with 70% EtOH/Heptaneat a flowrate of 1 mL/min, chiral purity=99.5%.

Example 139a and 139b: (S)-(S)-tetrahydrofuran-3-yl4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)pentanoateand (S)-(R)-tetrahydrofuran-3-yl4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)pentanoate

Example 139 (85 mg) was dissolved in Ethanol/Heptane (1:1, 1 mL) andpurified by chiral chromatography (stationary phase: Chiralpak IA,mobile phase: heptane/ethanol) (N28112-60). The combined fractionscontaining pure Isomer 1 (Example 139a) were concentrated in vacuo togive pure Isomer 1 as a colourless solid (25 mg). LCMS (System B):t_(RET)=0.92 min, MH⁺=537. The combined fractions containing pure Isomer2 (Example 139b) were concentrated in vacuo to give pure Isomer 2 as acolourless solid (45 mg). LCMS (System B): t_(RET)=0.93 min, MH⁺=537.

Example 162a and 162b: (2S,3R)-isobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(((R)-4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoateand (2S,3R)-isobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(((S)-4-methylmorpholin-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate

Example 162 (26 mg) was dissolved in Ethanol (1 mL) and purified bychiral chromatography (stationary phase: Chiralcel OD-H, mobile phase:60% heptane/40% ethanol) (N31661-32). The combined fractions containingpure Isomer 1 were concentrated in vacuo to give pure Isomer 1 (Example162a) as a colourless solid (11 mg). LCMS (System B): t_(RET)=0.88 min,MH⁺=540. The combined fractions containing pure Isomer 2 wereconcentrated in vacuo to give pure Isomer 2 (Example 162b) as acolourless solid (11 mg). LCMS (System B): t_(RET)=0.87 min, MH⁺=540.

Example 174a: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2-pyran-2-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate—Diastereomer1 and Example 174b: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate—Diastereomer2

Example 174 (203 mg) was dissolved in EtOH (2 mL) and purified by chiralchromatography (stationary phase Chiralpak AD-H, mobile phase 40%EtOH/Heptane).

The combined fractions were evaporated in vacuo to give purediastereomer 1 (Example 174a) as a colourless solid (81 mg). HPLC-UV: RT˜8.2 minutes, >99.5% isomeric purity by area HPLC @ 215 nm. LCMS (SystemI): t_(RET)=0.57 min, MH⁺=511. The combined fractions were evaporated invacuo to give pure diastereomer 2 (Example 174b) as a colourless solid(99 mg). HPLC-UV: RT ˜15.0 minutes, 99.3% isomeric purity by area HPLC @215 nm. LCMS (System I): t_(RET)=0.58 min, MH⁺=511.

Example 175: (2S,3R)-isobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate—Diastereoisomer2 and Example 175: (2S,3R)-isobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate—Diastereoisomer1

Example 175 (220 mg) was dissolved in EtOH (2 mL) and purified by chiralchormatography (stationary phase: Chiralpak AD-H, mobile phase: 40%EtOH/Heptane).

The combined fractions containing pure diastereomer 1 were concentratedin vacuo to give pure diastereomer 1 (Example 175a) as a colourlesssolid (91 mg). HPLC-UV: RT ˜8.3 minutes, >95% isomeric purity by areaHPLC @ 215 nm. LCMS (System I): t_(RET)=0.63 min, MH⁺=525. The combinedfractions containing pure diastereomer 2 were concentrated in vacuo togive pure diastereomer 2 (Example 175b) as a colourless solid (89 mg).HPLC-UV: RT ˜12.4 minutes, >95% isomeric purity by area HPLC @ 215 nm.

LCMS (System I): t_(RET)=0.63 min, MH⁺=525.

Example 178a: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate—Diastereoisomer1 and Example 178b: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydropyran-2n-yl)methyl)-1benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate—Diastereoisomer 2

Example 178 (100 mg) was dissolved in EtOH (2 mL) and purified by chiralchromatography (stationary phase: Chiralpak AD-H, mobile phase: 40%EtOH/Heptane). The combined fractions containing pure diastereomer 1were concentrated in vacuo to give pure diastereomer 1 as a colourlesssolid (50 mg). HPLC-UV: RT ˜8.1 minutes, >99.5% isomeric purity by areaHPLC @ 215 nm. LCMS (System I): t_(RET)=0.61 min, MH⁺=525. The combinedfractions containing pure diastereomer 2 were concentrated in vacuo togive pure diastereomer 2 as a colourless solid (41 mg). HPLC-UV: RT˜17.7 minutes, ca. >99.5% isomeric purity by area HPLC @ 215 nm. LCMS(System I): t_(RET)=0.61 min, MH⁺=525.

Example 180a: (2S,3R)-(S)-tetrahydrofuran-3-yl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate—Diastereomer 1and Example 180b: (2S,3R)-(S)-tetrahydrofuran-3-yl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate—Diastereomer2

Example 180 (140 mg) was dissolved in EtOH (2 mL) and purified by chiralchromatography (stationary phase: Chiralcel OD-H, mobile phase: 30%EtOH/Heptane). The combined fractions containing pure diastereomer 1(Example 180a) were concentrated in vacuo to give pure diastereomer 1 asa colourless solid (64 mg). HPLC-UV: RT ˜6.2 minutes, >99.5% isomericpurity by area HPLC @ 215 nm. LCMS (System I): t_(RET)=0.52 min,MH⁺=539. The combined fractions containing pure diastereomer 2 (Example180b) were concentrated in vacuo to give pure diastereomer 2 as acolourless solid (64 mg). HPLC-UV: RT ˜9.8 minutes, ca. >99.5% isomericpurity by area HPLC @ 215 nm. LCMS (System I): t_(RET)=0.52 min,MH⁺=539.

Example 181a: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate—Diastereomer1 and Example 181b: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate—Diastereomer2

Example 181 (107 mg) was dissolved in EtOH (2 mL) and purified by chiralchromatography (stationary phase: Chiralpak AD-H, mobile phase: EtOH).

The combined fractions containing pure Isomer 1 were concentrated invacuo to give pure Isomer 1 as a colourless solid (48 mg). HPLC-UV: RT˜8.4 minutes, >99.5% isomeric purity by area HPLC @215 nm. LCMS (SystemJ): t_(RET)=0.55 min, MH⁺=525. The combined fractions containing pureIsomer 2 were concentrated in vacuo to give pure Isomer 2 as acolourless solid (36 mg). HPLC-UV: RT ˜11.4 minutes, ca. 99.1% isomericpurity by area HPLC @ 215 nm. LCMS (System J): t_(RET)=0.56 min,MH⁺=525.

Example 182a: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate—Diastereomer1 and Example 182b: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-(tetrahydro-2H-pyran-4-yl)ethyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate—Diastereomer2

Example 182 (ca. 56 mg) was dissolved in EtOH (2 mL) and purified bychiral chormatography (stationary phase: Chiralpak IC, mobile phase:EtOH).

The combined fractions containing pure Isomer 1 were concentrated invacuo to give pure Isomer 1 as a colourless solid (27 mg). HPLC-UV: RT˜19.5 minutes, >99.5% isomeric purity by area HPLC @ 215 nm. LCMS(System J): t_(RET)=0.59 min, MH⁺=539. The combined fractions containingpure Isomer 2 were concentrated in vacuo to give pure Isomer 2 as acolourless solid (29 mg). HPLC-UV: RT ˜25.5 minutes, >99.5% isomericpurity by area HPLC @ 215 nm. LCMS (System J): t_(RET)=0.56 min,MH⁺=539.

Example 213a (2S,3R)-isobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(((R)-tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoateand 213b: (2S,3R)-isobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(((S)-tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate

A sample of (2S,3R)-isobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(For a preparation see Example 213, 125 mg) was purified by chiralchromatography on a Chiralpak IA (250×430 mm, 5 micron) column,Flowrate: 40 mL/min, detection: UV DAD (250 nm (bandwidth 80 nm,reference 375 nm (bandwidth 50 nm)). Eluting with a 50:50 mixture ofAcetonitrile containing 0.2% v/v isopropylamine and Propan-2-olcontaining 0.2% v/v isopropylamine. Appropriate fractions were combinedand evaporated under a stream of nitrogen to give the title compounds.

Isomer 1, Example 213a: (52 mg)

Chiral analysis carried out on a Chiralpak IA (250×4.6 mm, 5 micron)column, flowrate: 1 mL/min, detection: UV DAD (250 nm (bandwidth 40 nm,reference 375 nm (bandwidth 50 nm)). Eluting with a 50:50 mixture ofAcetonitrile containing 0.2% v/v isopropylamine and Propan-2-olcontaining 0.2% v/v isopropylamine. Chiral purity is >99.5%. LCMS(System J): t_(RET)=0.99 min; MH⁺525

Isomer 2, Example 213b: (53 mg)

Chiral analysis carried out on a Chiralpak IA (250×4.6 mm, 5 micron)column, flowrate: 1 mL/min, detection: UV DAD (250 nm (bandwidth 40 nm,reference 375 nm (bandwidth 50 nm)). Eluting with a 50:50 mixture ofAcetonitrile containing 0.2% v/v isopropylamine and Propan-2-olcontaining 0.2% v/v isopropylamine. Chiral purity is 99.7%. LCMS (SystemJ): t_(RET)=0.99 min; MH⁺525.

Example 225a and Example 225b: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(((R)-tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate“R-DIASTEREOISOMER” and (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(((S)-tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate“S-DIASTEREOISOMER”

The racemic (2R,3S)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(For a preparation see Example 225, 2.1 g) was separated by chiral HPLC.The HPLC purification was carried out on a Chiralpak ID column (Lot. No.ID12179-01, 250×30 mm). The purification was run using 80% EtOH incyclohexane with 0.2% IPA added, with a flow rate of 30 mL/min. UVdetection was at 215 nm. The first eluting enantiomer was collected, andthe fractions evaporated under reduced pressure to give the titlecompound “R-DIASTEREOISOMER” Example 225a (858 mg). The second elutingenantiomer was collected, and the fractions evaporated under reducedpressure, to give the title compound “S-DIASTEREOISOMER”, Example 225b(836 mg). DIASTEREOISOMER 1: LCMS (System J): t_(RET)=0.91 min; MH⁺ 511.DIASTEREOISOMER 2: LCMS (System J): t_(RET)=0.90 min; MH⁺ 511.

Example 228a and Example 228b: (2S,3R)-cyclobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoateDIASTEREOMER 1 and (2S,3R)-cyclobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoateDIASTEREOMER 2

The racemic (2S,3R)-cyclobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(For a preparation see Example 228, 250 mg) was separated by chiralHPLC. The HPLC purification was carried out on a Chiralpak AD-H column(Lot. No. ADH12143-01, 250×30 mm). The purification was run using 100%EtOH, with a flow rate of 25 mL/min. UV detection was at 215 nm. Thefirst eluting enantiomer was collected, and the fractions evaporatedunder reduced pressure. The sample was purified by MDAP (Method B). Thefractions were combined, evaporated under reduced pressure, and dried ina vacuum oven overnight to give the title compound “DIASTEREOISOMER 1”(98 mg). The second eluting enantiomer was collected, and the fractionsevaporated under reduced pressure, to give the title compound“DIASTEREOISOMER 2” (90 mg). DIASTEREOISOMER 1: LCMS (System J):t_(RET)=0.52 min; MH⁺509. DIASTEREOISOMER 2: LCMS (System J):t_(RET)=0.52 min; MH⁺509.

Example 229a and Example 229b: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoateDIASTEREOISOMER 1 and (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoateDIASTEREOISOMER 2

The racemic (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(for a preparation see Example 229, 200 mg) was separated by chiralHPLC. The HPLC purification was carried out on a Chiralpak AD-H column(Lot. No. ADH12143-01, 250×30 mm). The purification was run using 100%EtOH, with a flow rate of 25 mL/min. UV detection was at 215 nm. Thefirst eluting enantiomer was collected, and the fractions evaporatedunder reduced pressure to give the title compound “DIASTEREOISOMER 1”,Example 229a (21 mg). The second eluting enantiomer was collected, andthe fractions evaporated under reduced pressure, to give the titlecompound “DIASTEREOISOMER 2”, Example 229b (20 mg). DIASTEREOISOMER 1:LCMS (System J): t_(RET)=0.86 min; MH⁺ 497. DIASTEREOISOMER 2: LCMS(System J): t_(RET)=0.86 min; MH⁺ 497.

Example 232a and Example 232b: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoateDIASTEREOISOMER 1 and (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoateDIASTEREOISOMER 2

The racemic (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(For a preparation see Example 232, 80 mg) was separated by chiral HPLC.The HPLC purification was carried out on a Chiralpak AD-H column (Lot.No. ADH12143-01, 250×30 mm). The purification was run using 100% EtOH,with a flow rate of 25 mL/min. UV detection was at 215 nm. The firsteluting enantiomer was collected, and the fractions evaporated underreduced pressure to give the title compound “DIASTEREOISOMER 1”, Example232a (21 mg). The second eluting enantiomer was collected, and thefractions evaporated under reduced pressure, to give the title compound“DIASTEREOISOMER 2”, Example 232b (20 mg). DIASTEREOMER 1: LCMS (SystemJ): t_(RET)=0.92 min; MH⁺511. DIASTEREOMER 2: LCMS (System J):t_(RET)=0.92 min; MH⁺511.

Example 248a: (2S,3R)-isobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1((tetrahydro-2-pyran-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Enantiomer 1) and Example 248b: (2S,3R)-isobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1((tetrahydro-2-pyran-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Enantiomer 2)

The enantiomers were obtained by the chiral separation of Example 248.

Analytical Method:

-   -   Approx 0.5 mg dissolved in 50% EtOH/Heptane (1 mL) 20 uL        injected on column.    -   40% EtOH/Heptane, f=1.0 mL/min, wavelength 215 nm, 4. Ref        550,100    -   Column 4.6 mmid×25 cm Chiralcel OD-H    -   Lot No. ODHOCE-PD027

Prep Method:

-   -   Approx 110 mg dissolved in 1.5 mL of EtOH.    -   Injection; 1.5 mL the solution was injected onto the column.    -   40% EtOH/Heptane, f=30 mL/min, wavelength 215 nm, 4. Ref 550,100    -   Column 30 mm×25 cm Chiralcel OD-H    -   Lot No. ODH11158-01

Enantiomer 1 Rt=5.0 min. >99% ee by UV. Enantiomer 2 Rt=12.0 min. >99%ee by UV. Example 266a: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Enantiomer 1) and Example 266b: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Enantiomer 2)

The enantiomers were obtained by the chiral separation of Example 266.

Quantity: Approx 70 mg. Analytical Method:

-   -   Approx 0.5 mg dissolved in 50% EtOH/Heptane (1 mL) 20 ul        injected on column.    -   30% EtOH/Heptane, f=1.0 mL/min, wavelength 215 nm, 4. Ref        550,100    -   Column 4.6 mmid×25 cm Chiralcel OD-H    -   Lot No. ODHOCE-PD027

Prep Method:

-   -   Approx 70 mg dissolved in 1 mL EtOH.    -   Injection; 1 mL of the solution was injected onto the column.    -   30% EtOH/Heptane, f=30 mL/min, wavelength, 215 nm, 4. Ref        550,100    -   Column 30 mm×25 cm Chiralcel OD-H    -   Lot No ODH11158-01

Enantiomer 1 Rt=6.0 min. >99% ee by UV. Enantiomer 2 Rt=11.5 min. >99%ee by UV. Example 267a: (2S,3R)-isopropyl3-hydroxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoate(Enantiomer 1) and Example 267b: (2S,3R)-isopropyl3-hydroxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoate(Enantiomer 2)

The enantiomers were obtained by the chiral separation of Example 267

Quantity: Approx 120 mg. Analytical Method:

-   -   Approx 0.5 mg dissolved in 50% EtOH/Heptane (1 mL) 20 ul        injected on column.    -   20% EtOH/Heptane, f=1.0 mL/min, wavelength 215 nm, 4. Ref        550,100    -   Column 4.6 mmid×25 cm Chiralcel OD-H    -   Lot No. ODHOCE-PD027

Prep Method:

-   -   Approx 120 mg dissolved in 1 mL EtOH.    -   Injection; 1 mL of the solution was injected onto the column.    -   20% EtOH/Heptane, f=30 mL/min, wavelength, 215 nm, 4. Ref        550,100    -   Column 30 mm×25 cm Chiralcel OD-H    -   Lot No ODH11158-01

Enantiomer 1 Rt=8.5 min. >99% ee by UV. Enantiomer 2 Rt=12.2 min. >99%ee by UV. Example 270a: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Enantiomer 1) and Example 270b: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Enantiomer 2)

The enantiomers were obtained by the chiral separation of Example 270.

Quantity: Approx 150 mg. Analytical Method:

-   -   Approx 0.5 mg dissolved in 50% EtOH/Heptane (1 mL) 20 ul        injected on column.    -   30% EtOH/Heptane, f=1.0 mL/min, wavelength 215 nm, 4. Ref        550,100    -   Column 4.6 mmid×25 cm Chiralcel OD-H    -   Lot No. ODHOCE-PD027

Prep Method:

-   -   Approx 150 mg dissolved in 2 mL EtOH.    -   Injection; 1 mL of the solution was injected onto the column.    -   30% EtOH/Heptane, f=30 mL/min, wavelength, 215 nm, 4. Ref        550,100    -   Column 30 mm×25 cm Chiralcel OD-H    -   Lot No ODH11158-01

Enantiomer 1 Rt=6.0 min. >99% ee by UV. Enantiomer 2 Rt=11.5 min. >99%ee by UV. Example 271a: (2S,3R)-neopentyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1-H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre,Isomer 1) and Example 271b: (2S,3R)-neopentyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer2)

Example 271a: Isomer 1

Example 271b: Isomer 2

A sample of (rac)-(2S,3R)-neopentyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(for a preparation see Example 271, 280 mg) was separated by chiralcolumn chromatography using a 30 mm×25 cm Chiralcel OD-H column, elutingwith 50% EtOH/Heptane at a flowrate of 30 mL/min. The appropriatefractions for each isomer were combined and evaporated under reducedpressure to give the title compounds

Example 271a: Isomer 1 (140 mg)

LCMS (System I): t_(RET)=0.66 min; MH⁺ 513

Chiral analysis carried out on a 4.6 mmid×25 cm Chiralcel OD-H column,eluting with

50% EtOH/Heptane at a flowrate of 1 mL/min, chiral purity >99.5%

Example 271b: Isomer 2 (140 mg)

LCMS (System I): t_(RET)=0.65 min; MH⁺ 513

Chiral analysis carried out on a 4.6 mmid×25 cm Chiralcel OD-H column,eluting with

50% EtOH/Heptane at a flowrate of 1 mL/min, chiral purity >99.5%

Example 272a: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre,Isomer 1) and Example 272b: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer2)

Example 272a: Isomer 1

Example 272b: Isomer 2

A sample of (rac)-(2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(for a preparation see Example 272, 300 mg) was separated by chiralchromatography using a 30 mm×25 cm Chiralcel OD-H column, eluting with30% EtOH/Heptane at a flowrate of 30 mL/min. The appropriate fractionsfor each isomer were combined and evaporated under reduced pressure togive the title compounds

Example 272a: Isomer 1 (120 mg)

LCMS (System I): t_(RET)=0.58 min; MH⁺ 499

Chiral analysis carried out on a 4.6 mmid×25 cm Chiralcel OD-H column,eluting with

40% EtOH/Heptane at a flowrate of 1 mL/min, chiral purity >99.5%

Example 272b: Isomer 2 (120 mg)

LCMS (System I): t_(RET)=0.57 min; MH⁺ 499

Chiral analysis carried out on a 4.6 mmid×25 cm Chiralcel OD-H column,eluting with

40% EtOH/Heptane at a flowrate of 1 mL/min, chiral purity >99.5%

Example 273a: (2S,3R)-cyclobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre,Isomer 1) and Example 273b: (2S,3R)-cyclobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Single Diastereomer of Unknown Configuration at Marked Centre, Isomer2)

Example 273a: Isomer 1

Example 273b: Isomer 2

A sample of (rac)-(2S,3R)-cyclobutyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-methoxypropyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(for a preparation see Example 273, 290 mg) was separated by chiralchromatography using a 30 mm×25 cm Chiralcel OD-H column, eluting with40% EtOH/Heptane at a flowrate of 30 mL/min. The appropriate fractionsfor each isomer were combined and evaporated under reduced pressure togive the title compounds

Example 273a: Isomer 1 (120 mg)

LCMS (System I): t_(RET)=0.56 min; MH⁺ 497

Chiral analysis carried out on a 4.6 mmid×25 cm Chiralcel OD-H column,eluting with

40% EtOH/Heptane at a flowrate of 1 mL/min, chiral purity >99.5%

Example 273b: Isomer 2 (120 mg)

LCMS (System I): t_(RET)=0.57 min; MH⁺ 497

Chiral analysis carried out on a 4.6 mmid×25 cm Chiralcel OD-H column,eluting with

40% EtOH/Heptane at a flowrate of 1 mL/min, chiral purity >99.5%

Example 277: (S)-cyclopentyl2-((2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-6-yl)oxy)ethyl)amino)-3-methoxypropanoatehydrochloride

A mixture of2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-6-yl)oxy)acetaldehyde(intermediate 239, 116 mg, 0.229 mmol), (S)-cyclopentyl2-amino-3-methoxypropanoate hydrochloride (intermediate 196, 131 mg,0.587 mmol) and triethylamine (89 mg, 123 μL, 0.88 mmol) indichloromethane (5 mL) was stirred at room temperature for 30 minutes.Sodium triacetoxyborohydride (187 mg, 0.88 mmol) was added and thereaction mixture stirred at room temperature for 18 hours. SaturatedNaHCO₃ solution (15 mL) was added and the mixture stirred for 15minutes. The organic phase was separated and the aqueous phase wasextracted with dichloromethane (2×10 mL). The combined organics werewashed with brine, dried and evaporated. The residue was purified byMDAP (method B). The product was dissolved in ethyl acetate (5 mL) andtreated with 1.0M hydrogen chloride in diethyl ether (1.0 mL). Thesolvent was removed to give the title compound (53 mg, 0.088 mmol, 30.0%yield), as a colourless solid. LCMS (System B): t_(RET)=0.97 min; MH⁺567.

The following Examples were prepared in a similar manner to Example 277:

Example 278: S)-cyclopentyl 2-((2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H- benzo[d]imidazol-6-yl)oxy)ethyl)amino)propanoate hydrochloride (prepared from: Intermediate239 and Intermediate 61) System B, 0.98 min, MH⁺ = 537, Yield: 67 mg 40%

Example 279: (S)-(S)-tetrahydrofuran-3-yl 2-((2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-6-yl)oxy)ethyl)amino)-3- methylbutanoate hydrochloride(prepared from: Intermediate 239 and Intermediate 49) System B, 0.91min, MH⁺ = 567, Yield: 46 mg 26%

Example 280: (S)-(S)-1-methoxpropan-2-yl 2-((2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-6-yl)oxy)ethyl)amino)-3- methylbutanoate hydrochloride(prepared from: Intermediate 239 and Intermediate 63) System B, 1.01min, MH⁺ = 569, Yield: 58 mg 33%

Example 281a: (2S,3R)-isopropyl 3-hydroxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoate, hydrochloride(enantiomer 1) (prepared from: Intermediate 255 and Intermediate 31)System J, 0.81 min, MH⁺ = 483, Yield: 44 mg 36%

Example 281b: (2S,3R)-isopropyl 3-hydroxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoate hydrochloride (Enantiomer2) (prepared from: Intermediate 255 and Intermediate 31) System J, 0.81min, MH⁺ = 483, Yield: 53 mg 43%

Example 282a: (2S,3R)-isobutyl 3-hydroxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoate hydrochloride(enantiomer 1) (prepared from: Intermediate 251 and Intermediate 11)System J, 0.89 min, MH⁺ = 497, Yield: 42 mg 33%

Example 282b: (2S,3R)-isobutyl 3-hydroxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoate Hydrochloride (Enantiomer2) (prepared from: Intermediate 255 and Intermediate 11) System J, 0.89min, MH⁺ = 497, Yield: 30 mg 24%

Example 283a: (2S)-isobutyl 3-methoxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)propanoate hydrochloride(Enantiomer1) (prepared from: Intermediate 251 and Intermediate 66)System J, 0.94 min, MH⁺ = 497, Yield: 72 mg 57%

Example 283b: (2S)-isobutyl 3-methoxy-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydrofuran-3-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)propanoate hydrochloride (Enantiomer2) (prepared from: Intermediate 255 and Intermediate 66) System J, 0.94min, MH⁺ = 497, Yield: 48 mg 38%

Example 284: (S)-isopropyl 4-chloro-2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)butanoate hydrochloride (preparedfrom: Intermediate 116 and Intermediate 69) System J, 1.06 min, MH⁺ =529, (1 Cl) Yield: 280 mg 60%

Example 285: (2S)-tetrahydrofuran-3-yl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-methyl-1H-benzo[d]imidazol-5-yl)methyl)amino)- 3-methylbutanoatehydrochloride (prepared from: Intermediate 273 and Intermediate 39)System B, 0.88 min, MH⁺ = 453, Yield: 50 mg, 29%

Example 286: (S)-(S)-tetrahydrofuran-3-yl 4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)pentanoate (prepared from:Intermediate 115 and Intermediate 30) System B, 0.91 min, MH⁺ = 537,Yield: 40 mg, 44%

Example 287: (2S,3R)-isobutyl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoate (prepared from:Intermediate 116 and Intermediate 11) System C, 0.60 min, MH⁺ = 525,Yield: 80 mg, 40%

Example 288: (2S,3R)-isobutyl2-((2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2-pyran-4-yl)methyl)-1-benzo[d]imidazol-6-yl)oxy)ethyl)amino)-3-hydroxybutanoate,Hydrochloride

Stage i)

A mixture of2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-6-yl)oxy)acetaldehyde(Intermediate 239) (116 mg, 0.293 mmol), (2S,3R)-isobutyl2-amino-3-hydroxybutanoate 4-methylbenzenesulphonic acid salt(Intermediate 11) (204 mg, 0.587 mmol) and triethylamine (89 mg, 123 μL,0.88 mmol) in dichloromethane (5 mL) was stirred at room temperature for30 minutes. Sodium triacetoxyborohydride (187 mg, 0.88 mmol) was addedand the reaction mixture stirred at room temperature for 18 hours.Saturated NaHCO₃ solution (15 mL) was added and the mixture stirred for15 minutes. The organic phase was separated and the aqueous phase wasextracted with dichloromethane (2×10 mL). The combined organics werewashed with brine, dried and evaporated to give stage i) product. LCMSMass ion [M−2] with respect to expected observed. Suggests enamine hasformed.

Stage ii)

The stage i) product was dissolved in isopropanol (10 mL). The solutionwas treated with ammonium formate (92 mg, 1.47 mmol) and 10% palladiumon carbon, 50% water paste (23 mg, 20% wt). The reaction mixture wasrefluxed for 4 hours. The cooled reaction mixture was filtered through‘celite’. The solvent was evaporated from the filtrate and the residuepurified by high pH MDAP (Method B). Not pure enough, repurified byformic acid MDAP (Method A). The product was dissolved in ethyl acetate(5 mL) and treated with 1.0M hydrogen chloride in diethyl ether (1 mL).The solvent was removed to give the title compound (2 mg, 3.38 μmol,1.153% yield), as a colourless solid. LCMS (System B): t_(RET)=0.92 min;MH⁺ 555.

The following Example was prepared in a similar manner to Example 288:

Example 289: (2S,3R)-isopropyl 2-((2-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-6-yl)oxy)ethyl)amino)-3- hydroxybutanoate hydrochloride(prepared from: Intermediate 239 and Intermediate 31) System B, 0.84min, MH⁺ = 541, Yield: 4 mg, 2.3%

Example 290: (2S,3R)-isopropyl2-((2-(2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)ethyl)amino)-3-hydroxybutanoate

1,3-dimethyl-5-(5-(oxiran-2-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)pyridin-2(1H)-one(for a preparation see Intermediate 266, 0.1 g, 0.264 mmol) wasdissolved in THF (3 mL) under nitrogen. The solution was cooled down to0° C. and BF₃.OEt₂ (0.017 mL, 0.132 mmol) was added. The mixture wasstirred at 0° C. for 5 min, then (2S,3R)-isopropyl2-amino-3-hydroxybutanoate hydrochloride (Intermediate 31a, 0.057 g,0.290 mmol), triethylamine (0.110 mL, 0.791 mmol) and p-toluenesulfonicacid monohydrate (0.050 g, 0.264 mmol) were added. The reaction mixturewas stirred under nitrogen at room temperature for 1 hr. Sodiumtriacetoxyborohydride (0.223 g, 1.054 mmol) was added. The reactionmixture was stirred under nitrogen for 2 hours. The reaction mixture wasstirred 10 min with a saturated solution of sodium bicarbonate, it wasthen diluted with ethyl acetate. The layers were separated. The aqueouswas extracted with ethyl acetate. The organics were combined and driedover a phase separator, and concentrated. The oil was purified by flashcolumn chromatography eluted with 0-10% MeOH in DCM. The fractions wereconcentrated to dryness. The residue was purified by MDAP (method B).The solvent was evaporated in vacuo to give the title compound as awhite solid (40% yield). LCMS (System J): t_(RET)=0.91 min; MH⁺ 525.

The following Example was prepared in a similar manner to Example 290:

Example 291: (2S,3R)-tert-butyl 2-((2-(2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)ethyl)amino)-3- hydroxybutanoate (prepared from:Intermediate 266 and (2S,3R)-tert-butyl 2-amino-3- hydroxybutanoatehydrochloride (commercially available)) System J, 0.96 min, MH⁺ = 539,Yield: 23 mg, 11%

Example 292: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoatehydrochloride

(2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(Example 303, 400 mg, 0.788 mmol) was dissolved in ethyl acetate (3 mL)and treated with 1.0M hydrogen chloride in diethyl ether (1 mL). Diethylether (20 mL) was added, the solid was filtered off, washed with diethylether and dried to give the title compound (409 mg, 0.748 mmol), as acolourless solid. LCMS (System I): t_(RET)=0.51 min; MH⁺ 511. ¹H NMR(400 MHz, DMSO-d6) ⋅ ppm 9.78 (br s, 2H), 8.44 (s, 1H), 8.02-8.10 (m,1H), 7.83 (s, 1H), 7.67 (d, J=12.0 Hz, 1H), 4.88-4.97 (m, 1H), 4.38-4.49(m, 4H), 4.08-4.14 (m, 1H), 3.70-3.77 (m, 2H), 3.61-3.67 (m, 1H), 3.58(s, 3H), 3.07-3.16 (m, 2H), 2.13 (s, 3H), 1.93-2.06 (m, 1H), 1.15-1.32(m, 13H).

Example 293:(2S,3R)-2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoicacid, Hydrochloride

(2S,3R)-2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoicacid (Example 82, 8 mg, 0.017 mmol) was triturated with 1M hydrogenchloride in diethyl ether (17 mL, 0.017 mmol) and stirred under nitrogenfor 5 hours. Solvents were then removed in vacuo, the white solidobtained was dried in a vacuum oven at 40° C. for 24 hours to give thetitle compound (5.6 mg, 0.011 mmol, 64.9% yield) and appeared as a whitesolid. LCMS (System I): t_(RET)=0.51 min; MH⁺ 469.

Example 294: (2S,3R)-cyclobutyl2-(((1-(((S)-1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoatehydrochloride

(2S,3R)-cyclobutyl 2-amino-3-hydroxybutanoate, 4-Methylbenzenesulphonicacid salt (for a preparation see intermediate 32, 170 mg, 0.492 mmol)was added to a solution of(S)-1-((1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-benzo[d]imidazole-5-carbaldehyde(Intermediate 135) (100 mg, 0.246 mmol) in DCM (5 mL) and the resultingsolution stirred overnight under N₂. Sodium triacetoxyborohydride (156mg, 0.738 mmol) was added and the resulting suspensions stirred for 3 h.The reaction mixture was solubilised with MeOH and loaded on to a 10 gSCX cartridge. The cartridge was eluted with MeOH, followed by 2Mmethanolic ammonia. The basic fractions were evaporated in vacuo todryness and the residues purified by MDAP (HPH method B). The productcontaining fractions were evaporated in vacuo azeotroping with EtOH togive a colourless oil. The residues were dissolved in DCM (1 mL) and 1 MHCl in diethyl ether added with stirring. The resulting suspension wasevaporated in vacuo to give the title compound as a white solid. Thetotal yield of the reaction was 53%.

LCMS (System C): t_(RET)=0.52 min, MH⁺=564

The following Examples were prepared in a similar manner to Example 294:

Example 295: (2S,3R)-tert-butyl 2-(((1-(((S)-1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate, Hydrochloride (prepared from:Intermediate 135 and (2S,3R)-tert-butyl 2-amino-3- hydroxybutanoatehydrochloride (commercially available)) System C, 0.54 min, MH⁺ = 566,Yield: 69 mg, 42%

Example 296: (2S,3R)-isopropyl 2-(((1-(((S)-1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate, Hydrochloride (prepared from:Intermediate 135 and Intermediate 31) System C, 0.50, MH⁺ = 552, Yield:20 mg, 13%

Example 297: (2S,3R)-isopropyl 2-(((1-(((R)-1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate, Hydrochloride (prepared from:Intermediate 136 and Intermediate 31) System I, 0.51 min, MH⁺ = 552,Yield: 78 mg, 48%

Example 298: (2S,3R)-cyclobutyl 2-(((1-(((R)-1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate, Hydrochloride (prepared from:Intermediate 136 and Intermediate 32) System I, 0.54 min, MH⁺ = 564,Yield: 34 mg, 22%

Example 299: (S)-isopropyl 2-(((1-(((S)-1-acetylpiperidin-3-yl)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-methylbutanoate, Hydrochloride (prepared from:Intermediate 135 and Intermediate Intermediate 12) System C, 0.56 min,MH⁺ = 550, Yield: 126 mg, 79%

Example 300: (S)-cyclopentyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-methyl-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-methoxypropanoate,hydrochloride

2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-methyl-1h-benzo[d]imidazole-5-carbaldehyde(102 mg, 0.363 mmol, Intermediate 273) was dissolved in DCM (5 mL) andtreated with (S)-cyclopentyl 2-amino-3-methoxypropanoate, hydrochloride(162 mg, 0.725 mmol, Intermediate 196) and triethylamine (0.151 mL,1.088 mmol). To the stirred solution was added sodiumtriacetoxyborohydride (384 mg, 1.813 mmol). The mixture was then stirredfor 18 hrs under nitrogen. Saturated sodium bicarbonate (15 mL) was thenadded to the mixture and stirred for 15 min. The layers were thenseparated, the aqueous layer extracted twice with dichloromethane (10mL×2) and the organics combined and dried with a hydrophobic frit. Thesolvent was then removed under vacuum. The residue was dissolved inminimum amount of dichloromethane and loaded onto a 10 g silicacartridge and eluted using the following method: 0% ethyl acetate for 2column volumes, 0-25% ethanol/ethyl acetate for 10 column volumes then25% ethanol/ethyl acetate for 5 column volumes. The appropriatefractions were combined and the solvent removed in vacuo producing(S)-cyclopentyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-methyl-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-methoxypropanoateas a colourless oil. The sample was dissolved in a minimum amount ofdiethyl ether and then treated with a few drops of 1M hydrochloric acidin diethyl ether and the solvent and HCl removed under vacuum. Thisyielded the title compound (68 mg, 0.139 mmol, 38.4% yield) as a whitesolid. LCMS (System B): t_(RET)=0.94 min, MH⁺=453.

Example 301: (S)-cyclopentyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-methyl-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxy-3-methylbutanoate, hydrochloride

2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-methyl-1H-benzo[d]imidazole-5-carbaldehyde(100 mg, 0.355 mmol, Intermediate 273) was dissolved in DCM (5 mL) andtreated with (S)-cyclopentyl 2-amino-3-hydroxy-3-methylbutanoate,hydrochloride (110 mg, 0.463 mmol, Intermediate 51) and triethylamine(0.148 mL, 1.066 mmol) and stirred for 2 hrs. Sodiumtriacetoxyborohydride (377 mg, 1.777 mmol) was then added and thereaction stirred overnight. After this time, another 3 equivalents ofsodium triacetoxyborohydride (377 mg, 1.777 mmol) was added and themixture stirred for 2 hrs. To the mixture was added sodium bicarbonate(25 mL) and the reaction stirred for 15 min. The phases were thenseparated and the aqueous layer extracted twice with dichloromethane(2×10 mL) and the organics combined and dried using a hydrophobic frit.The solvent was then removed under vacuum and the residue dissolved inminimum of dichloromethane. The solution was then loaded onto a BiotageSNAP 10 g cartridge and automated chromatography run using the followingmethod: 0% ethyl acetate for 2 column volumes, 0-25% ethanol in ethylacetate over 10 column volumes, 25% ethanol for 5 column volumes. Theappropriate fractions were then combined and solvent removed undervacuum. The residue was then dissolved in a minimum of dichloromethaneand then loaded onto a Biotage SNAP 10 g cartridge and automatedchromatography run using the following method: 15% ethanol in ethylacetate for 2 column volumes, 15-40% ethanol in ethyl acetate for 10column volumes, 40% ethanol in ethyl acetate for 5 column volumes. Theresidue was then dissolved in dimethyl sulphoxide (0.4 mL) anddichloromethane (0.4 mL) and purified by high pH mass directedautopurification. The appropriate fractions were then combined and thesolvent removed under vacuum. This yielded (S)-cyclopentyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-methyl-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxy-3-methylbutanoate(26.5 mg, 0.057 mmol, 15.98% yield) as a yellow oil. The oil was thendissolved in diethyl ether and a few drops of 1M HCl in diethyl etheradded and the solvent removed under vacuum yielding the title compound(26.5 mg, 0.053 mmol, 14.82% yield) as a white solid. LCMS (System B):t_(RET)=0.95 min, MH⁺=467.

Example 302:(S)-4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methylpiperidin-4-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)pentanoicacid, hydrochloride

To a solution of (S)-cyclopentyl4-methyl-2-(((2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(1-methylpiperidin-4-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)pentanoate(For a preparation see Example 2, 17 mg, 0.032 mmol) in Methanol (1 mL)and THF (1 mL) was added 1 M aqueous lithium hydroxide solution (0.032mL, 0.032 mmol) and the reaction mixture stirred at room temperatureovernight. Stirring was continued for three days at room temperature.The reaction mixture was heated at 50° C. for two days. Further 1 Maqueous lithium hydroxide solution (0.16 mL, 0.16 mmol) was added, andthe reaction mixture heated at 50° C. overnight. The sample wasevaporated under reduced pressure, and the residue dissolved in water (5mL) and loaded onto an isolute 103 cartridge. The cartridge was washedwith water, and then eluted with methanol. The methanol fractions werecombined and blown down under a stream of nitrogen to give the titlecompound (13 mg). LCMS (System B): t_(RET)=0.57 min; MH⁺ 466.

Example 303: (2S,3R)-Isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate

A mixture of2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(for an example preparation see Intermediate 116, 10.0 g, 27.4 mmol),(2S,3R)-isopropyl 2-amino-3-hydroxybutanoate hydrochloride (for anexample preparation see Intermediate 31a, 10.82 g, 54.7 mmol) andtriethylamine (6.92 g, 9.54 mL, 68.4 mmol) in DCM (100 mL) was stirredat room temperature overnight. Sodium triacetoxyborohydride (17.4 g, 82mmol) was added and the reaction mixture was stirred at room temperaturefor 4 hours. The reaction mixture was diluted with DCM (100 mL) andpoured onto saturated sodium bicarbonate (300 mL). The mixture wasstirred at room temperature for 30 minutes, the organic phase wasseparated and the aqueous phase was extracted with DCM (2×100 mL). Thecombined organics were dried and evaporated. The residue waschromatographed using a gradient of 2.5-10% MeOH in DCM to give thetitle compound (11.57 g, 22.66 mmol, 83% yield), as a colourless foam.LCMS (System J): t_(RET)=0.87 min; MH⁺ 511. ¹H NMR (d₆-DMSO): δ1.06-1.26 (m, 13H), 1.90-2.02 (m, 1H), 2.11 (s, 3H), 2.20-2.41 (m, 1H),3.00 (d, J=4.9 Hz, 1H), 3.05-3.16 (m, 2H), 3.56 (s, 3H), 3.66 (d, J=1.32Hz, 1H), 3.76-3.86 (m, 1H), 3.90 (d, J=13.2 Hz, 1H), 4.26 (d, J=7.3 Hz,2H), 4.67 (d, J=5.4 Hz, 1H), 4.93 (sept, J=6.2 Hz, 1H), 7.22 (dd, J=8.3,1.2 Hz, 1H), 7.53 (s, 1H), 7.61 (d, J=8.3 Hz, 1H), 7.73 (dd, J=2.3, 1.1Hz, 1H), 8.12 (d, J=2.2 Hz, 1H).

Alternative Process for the Preparation of Example 303:(2S,3R)-Isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate

(3-Nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)methanol

(4-fluoro-3-nitrophenyl)methanol (2.4 g, 14.02 mmol) and(tetrahydro-2H-pyran-4-yl)methanamine (2.423 g, 21.04 mmol) weresuspended in water (30 ml) and potassium carbonate (2.52 g, 18.23 mmol)was added, then the mixture was stirred at 80° C. for 24h, then allowedto cool while stirring. The resulting mixture was extracted with EtOAc(50 ml) and the organic layer washed with water (50 ml), dried andevaporated in vacuo to give the title compound (3.60 g, 13.52 mmol, 96%yield) as a dark yellow solid. LCMS (System D): t_(RET)=0.82 min; MH⁺267. The title compound was used in the next step without purification.

(3-Amino-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)methanol

(3-Nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)methanol (48g, 180 mmol) was dissolved in ethanol (400 ml) and hydrogenated overPd/C 5% by weight (3 g, 28.2 mmol) at atmospheric pressure for 18h, thenthe mixture was filtered through Celite under nitrogen, and the filtrateevaporated in vacuo to give the title compound (50 g, 212 mmol, 117%yield) as a dark brown oil. LCMS (System D): t_(RET)=0.62 min; MH⁺ 237.Product was carried through to the next step without furtherpurification.

5-(5-(Hydroxymethyl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2-(1H)-one

(3-Amino-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)methanol (50g, 190 mmol) was suspended in water (500 ml) and1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carbaldehyde (31.7 g, 209 mmol)and cetylpyridinium bromide (14.64 g, 38.1 mmol) were added, then themixture was stirred vigorously overnight. The mixture was extracted withDCM (3×300 ml) and the combined organics were washed with brine (500ml), then dried and evaporated to give a dark brown solid. This wassuspended in EtOAc (500 ml) and heated to reflux for 2h, then cooled andthe product collected by filtration. The crude was resuspended in EtOAc(500 ml) and heated to reflux again, then cooled in an ice bath and theproduct collected by filtration and washed with ether (300 ml) to give abrown solid (64 g). LCMS showed clean product, but the NMR spectrumshows the presence of 0.2 eq of the cetylpyridinium salt remaining inthe product. Carried through to the next step without purification. Thetitle compound was 90% Wt purity. LCMS (System D): t_(RET)=0.66 min; MH⁺368.

(2S,3R)-isopropyl 2-amino-3-hydroxybutanoate, hydrochloride

AcCl (96 ml, 1343 mmol) was added dropwise to 2-propanol (500 ml, 6490mmol) and the mixture was then stirred for 20 min before addition of(2S,3R)-2-amino-3-hydroxybutanoic acid (40 g, 336 mmol). The resultingsuspension was heated to reflux overnight, then cooled and evaporated invacuo to give a colourless oil. This was triturated with ether (300 ml)and the product collected by filtration to give (2S,3R)-isopropyl2-amino-3-hydroxybutanoate as a colourless solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.49 (br. s., 3H), 5.66 (br. s., 1H), 4.99(td, J=6.24, 12.47 Hz, 1H), 4.09 (br. s., 1H), 3.80 (d, J=4.16 Hz, 1H),1.17-1.29 (m, 9H)

2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde

5-(5-(Hydroxymethyl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(64 g, 122 mmol) was dissolved in DCM (600 ml) and manganese dioxide(42.4 g, 488 mmol) was added, then the mixture was heated at reflux for18h. LCMS showed complete conversion, and the mixture was filtered andthe solid washed with DCM. The filtrate was evaporated in vacuo to givea brown gum, which was dissolved in DCM (100 ml) and loaded onto a 340 gsilica column, then eluted with 0-50% EtOH/EtOAc and product-containingfractions were evaporated in vacuo to give a brown solid. This wastriturated with ether (200 ml) and the solid collected by filtration,then suspended in EtOAc (300 ml) and heated to reflux for 1h, thencooled in an ice bath and the product collected by filtration to give2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(42.5 g, 116 mmol, 95% yield) as a sand-coloured solid. LCMS (System D):t_(RET)=0.74 min; MH⁺ 366. The filtrate was evaporated in vacuo and theresidue triturated in EtOAc (50 ml) at reflux for 30 min, then cooledand filtered to give an additional portion of the product (3 g) as abeige solid, NMR consistent with the desired aldehyde. The titlecompound was 80% Wt purity.

(2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate

2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazole-5-carbaldehyde(42 g, 115 mmol) and (2S,3R)-isopropyl 2-amino-3-hydroxybutanoatehydrochloride (34.1 g, 172 mmol) were dissolved in DCM (500 ml), thenEt₃N (48.1 ml, 345 mmol) was added, followed by sodiumtriacetoxyborohydride (73.1 g, 345 mmol) and the mixture was stirred atroom temperature for 24h. The mixture was added to 1.5 litres ofsaturated sodium bicarbonate solution in a 5 litre conical flask andstirred vigorously for 1h, then the organic layer was separated, theaqueous extracted with DCM (500 ml) and the combined organics washedwith water (500 ml) and brine (500 ml), dried over sodium sulphate andevaporated in vacuo to give a brown foam. The crude product wasdissolved in DCM (200 ml) and loaded onto a 750 g silica column, theneluted with 0-30% EtOH/EtOAc and clean product-containing fractions wereevaporated in vacuo to give (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(51 g, 100 mmol, 87% yield) as a beige foam. LCMS (System D):t_(RET)=0.86 min; MH⁺ 511.

¹H NMR (d6-DMSO): δ 1.06-1.26 (m, 13H), 1.90-2.02 (m, 1H), 2.11 (s, 3H),2.20-2.41 (m, 1H), 3.00 (d, J=4.9 Hz, 1H), 3.05-3.16 (m, 2H), 3.56 (s,3H), 3.66 (d, J=1.32 Hz, 1H), 3.76-3.86 (m, 1H), 3.90 (d, J=13.2 Hz,1H), 4.26 (d, J=7.3 Hz, 2H), 4.67 (d, J=5.4 Hz, 1H), 4.93 (sept, J=6.2Hz, 1H), 7.22 (dd, J=8.3, 1.2 Hz, 1H), 7.53 (s, 1H), 7.61 (d, J=8.3 Hz,1H), 7.73 (dd, J=2.3, 1.1 Hz, 1H), 8.12 (d, J=2.2 Hz, 1H).

Preparation of the Edisylate Salt of Example 303 (Without Seeding):(2S,3R)-Isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,1,2-ethanedisulphonic acid salt

To a carousel tube was added (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(For a preparation see Example 303, 100 mg, 0.196 mmol) and isopropanol(1.35 mL). The stirred mixture was heated to 40° C. and a solution ofethane-1,2-disulfonic acid (44.7 mg, 0.235 mmol) in isopropanol (557 μL)was added. The mixture was stirred at 40° C. for 15 hr. After this time,solid had formed. The reaction was removed from the carousel, cooleddirectly to 24° C. and stirred for 6 h. After this time, the suspensionwas filtered and dried under vacuum for 5 min. The solid, transferredinto a vial was further dried in the vacuum oven at 40° C. for 3 days toyield (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,1,2-ethanedisulphonic acid salt (91 mg, 66.3% yield) as a whitecrystalline solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.61 (1H, br. s.),9.29 (1H, br. s.), 8.35 (1H, br. s.), 8.02 (1H, d, J=8.6 Hz), 7.92 (1H,s), 7.82-7.75 (1H, m), 7.58 (1H, d, J=8.6 Hz), 4.94 (1H, spt, J=6.2 Hz),4.46-4.34 (4H, m), 4.05 (1H, quin, J=6.4 Hz), 3.77-3.69 (2H, m), 3.66(1H, br. s.), 3.59 (3H, s), 3.17-3.08 (2H, m), 2.66 (4H, s), 2.13 (3H,s), 2.06-1.93 (1H, m), 1.31-1.16 (13H, m). LCMS (System D): t_(RET)=0.88min; MH⁺ 511.

Preparation of the Edisylate Salt of Example 303 (with Seeding):(2S,3R)-Isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate,1,2-ethanedisulphonic acid salt

To an EasyMax 400 mL reactor was added (2S,3R)-Isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(for an example preparation see Example 303 (alternative process))(10 g,19.58 mmol) and Isopropanol (135 mL). In a separate flask was prepared asolution of ethane-1,2-disulfonic acid (4.47 g, 23.50 mmol) inIsopropanol (28 mL) warmed at 40° C. and filtered.

To the solution of (2S,3R)-Isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(for an example preparation see Example 303 (alternative process),stirred at 250 rpm was added 40% of the ethane-1,2-disulfonic acidsolution (11.2 mL). A seed of (2S,3R)-Isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(for an example preparation see above ‘Preparation of the edisylate saltof Example 303 (without seeding)’)) (131 mg) was added and the mixturewas stirred at 40° C. for 1.5 h. After this time, the remaining 60% ofthe ethane-1,2-disulfonic acid solution (16.8 mL) was added dropwiseover 6 h. On complete addition of ethane-1,2-disulfonic acid solutionthe mixture was slowly cooled to 20° C. over 3.5 h and stirred for afurther 11 h at RT. The resulting suspension was filtered with filtercup and paper filter, with the filtrate running clear. The filter cakewas washed with IPA (2×20 mL and 10 mL) and further dried under withvacuum to yield the wet filter cake (24.06 g). The solids were collectedand dried in a vacuum oven (44° C.) for 22 h to yield the title compound(11.337 g, 16.02 mmol, 82% yield) as a white crystalline solid. LCMS(System D): t_(RET)=0.88 min; MH⁺ 511.

Example 304:(2S,3R)-2-(((2-(1,5-Dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(((S-tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoicacid

(2S,3R)-(S)-tetrahydrofuran-3-yl-2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(((S)-tetrahydrofuran-2-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate(For a preparation see Example 224, 190 mg, 0.362 mmol) was dissolved inTHF (3 mL) and methanol (3 mL). Lithium hydroxide (26.0 mg, 1.087 mmol)was dissolved in water (1 mL) and the solution was added. The reactionmixture was stirred for 5 hours at 40° C. 2M hydrochloric acid (0.543mL, 1.087 mmol) was added and the solvents were removed under reducedpressure. The residue was dissolved in water (1 mL) and DMSO (0.8 mL),the solution was filtered, and purified by MDAP (Method B). Theappropriate fractions were combined and concentrated under reducedpressure. The residue was suspended in THF (3 mL) and 2 M HCl (0.2 mL)was added. The solvents were removed under a stream of nitrogen. Thesample was dissolved in water (0.7 mL) and DMSO (0.7 mL) and purified byMDAP (Method B). The appropriate fractions were blown under a stream ofnitrogen and the resultant residues were treated with THF (4 mL) and 2 Msodium hydroxide (0.04 mL). The solvents were blown down under a streamof nitrogen. The sample was dissolved in ethyl acetate, filtered, andthe solid which was isolated during filtration was dissolved in waterand concentrated under reduced pressure. The residue was dissolved inwater (0.8 mL) and purified by MDAP (Method B). The appropriatefractions were blown under a stream of nitrogen to yield the titlecompound (16 mg) as a white solid. LCMS (System J): t_(RET)=0.54 min;MH⁺ 455.

Example 305: (2S,3R)-Isopropyl2-(((1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-yl)methyl)amino)-3-hydroxybutanoate

To a mixture of1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazole-5-carbaldehyde(for an example preparation see Intermediate 224, 6.3 g, 17.05 mmol) and(2S,3R)-isopropyl 2-amino-3-hydroxybutanoate hydrochloride (for anexample preparation see Intermediate 31a, 6.74 g, 34.1 mmol) inDichloromethane (DCM) (200 mL) was added triethylamine (5.94 mL, 42.6mmol) and the reaction mixture stirred at room temperature overnight.Sodium triacetoxyborohydride (10.84 g, 51.2 mmol) was added and thereaction mixture stirred at room temperature for four hours. Saturatedaqueous sodium bicarbonate solution was added (200 mL) and the organiclayer separated. The aqueous layer was re-extracted with dichloromethane(2×100 mL), and the organic layers combined, dried using a hydrophobicfrit and evaporated under reduced pressure. The sample was loaded indichloromethane and purified by column chromatography (silica) using agradient of 0-5% methanol in dichloromethane. The appropriate fractionswere combined and evaporated under reduced pressure. The sample waspurified by HPLC (Xselect CSH C18 column, eluted usingwater/acetonitrile with 0.1% formic acid modifier) and the fractionsevaporated under reduced pressure to give isolated material (5.7 g). 2.1g of material was removed for subsequent reactions. The remainingmaterial (3.6 g), a colourless gum, was dried in a vacuum oven for 3nights whereupon it became a scratchable glass. As much material aspossible was removed from the flask and returned to a vacuum oven for afurther 2 nights to give the title compound (3.2 g, 6.22 mmol). LCMS(System J): t_(RET)=0.92 min; MH⁺ 515. ¹H NMR (d₆-DMSO): δ 1.11 (d,J=6.4 Hz, 3H), 1.19 (d, J=6.1 Hz, 6H), 2.09 (s, 3H), 3.00 (d, J=4.9 Hz,1H), 3.16 (d, J=1.5 Hz, 6H), 3.54 (s, 3H), 3.65 (d, J=13.2 Hz, 1H),3.70-3.86 (m, 3H), 3.91 (d, J=13.0 Hz, 1H), 3.96-4.07 (m, 2H), 4.70 (d,J=5.4 Hz, 1H), 4.78-4.86 (m, 1H), 4.94 (sept, J=6.2 Hz, 1H), 7.18 (dd,J=8.4, 1.3 Hz, 1H), 7.55 (s, 1H), 7.67-7.70 (s, 1H), 7.72 (d, J=8.6 Hz,1H), 8.05 (d, J=2.2 Hz, 1H).

Example 306: (2S,3S)-(S)-tetrahydrofuran-3-yl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate

To a stirred solution of2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[o]imidazole-5-carbaldehyde(0.438 mmol, 160 mg) (for an example preparation see Intermediate 116)and ((2S,3S)-(S)-tetrahydrofuran-3-yl 2-amino-3-hydroxybutanoatehydrochloride (98.8 mg, 0.525 mmol) (for an example preparation seeIntermediate 40) in DCM (5 mL) was added triethylamine (0.182 mL). Theresulting mixture was stirred for 1 h and sodium triacetoxyborohydride(278 mg, 1.374 mmol) was added. The reaction mixture was stirred for 30min and allowed to stand overnight The reaction mixture was partitionedbetween DCM and aq. sat. NaHCO₃. The organic layer was removed and theaqueous extracted twice with DCM. The organic phases were combined andconcentrated in vacuo to give a brown oil. The oil was dissolved in DCMand purified by silica gel chromatography eluting with EtOAc:EtOH(3-25%). The appropriate fractions were combined and evaporated in vacuoto give the title compound as a colourless oil. The total yield of thereaction was 46%. LCMS (System C): t_(R)ET=0.45 min, MH⁺=539.

Example 307: (2S,3R)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((S)-piperidin-3-ylmethyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate

5-6N HCl in isopropanol (0.2 mL) was added to a solution of(R)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-((((2S,3R)-3-hydroxy-1-isopropoxy-1-oxobutan-2-yl)amino)methyl)-1-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate(For a preparation see Intermediate 267, 31.4 mg, 51.5 mmol), inisopropanol (1 mL). Resulting mixture was stirred at r.t. overnight (17hrs). 5-6N HCl in isopropanol (0.8 mL) was added to reaction mixture andresulting solution was stirred for 5 hours. Volatiles were removed undera stream of nitrogen and mixture was purified by MDAP (Method B)Fractions were combined, volatiles removed under reduce pressure andcrude was loaded on a 2 g pre equilibrated SCX cartridge, eluted with3CV of methanol and 3CV of 2M ammonia in Methanol. Basic fractions werecombined and volatiles were removed under reduced pressure to afford ayellow oil. The oil was purified by MDAP (Method B). Relevant fractionscombined and volatiles were removed under reduce pressure to give thetitle compound (15.5 mg) as a translucent/white solid.

LCMS (System J): t_(RET)=0.79 min; MH⁺ 510

Example 308: (2S,3R)-tert-butyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((S)-piperidin-3-ylmethyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-hydroxybutanoate

5-6N HCl in isopropanol (0.2 mL) was added to a solution of(R)-tert-butyl3-((6-((((2S,3R)-1-(tert-butoxy)-3-hydroxy-1-oxobutan-2-yl)amino)methyl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate(For a preparation see Intermediate 268, 21 mg, 33.7 mmol) inisopropanol (1 mL). Resulting mixture was stirred at r.t. overnight (17hrs). 5-6N HCl in isopropanol (0.8 mL) was added to reaction mixture andresulting solution was stirred for 5 hours. Volatiles were removed usingblow down unit and mixture was purified by MDAP (Method B). Fractionswere combined, volatiles removed under reduced pressure and crude wasloaded on a 2 g pre equilibrated SCX cartridge, eluted with 3CV ofmethanol and 3CV of 2M ammonia in Methanol. Basic fractions werecombined and volatiles were removed under reduce pressure to afford ayellow oil. This oil was purified by MDAP (Method B). Relevant fractionscombined and volatiles were removed under reduce pressure to afford thetitle compound (13.5 mg) as a translucent/white solid. LCMS (System J):t_(RET)=0.85 min; MH⁺ 524.

Example 309: (S)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((R)-piperidin-3-ylmethyl)-1H-benzo[d]imidazol-6-yl)methyl)amino)-3-methylbutanoatedihydrochloride

HCl 5M in IPA (1 mL, 5.00 mmol) was added to (S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-6-((((S)-1-isopropoxy-3-methyl-1-oxobutan-2-yl)amino)methyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate(For a preparation see Intermediate 269, 56 mg, 0.092 mmol) and theresulting solution was stirred at rt for 21 hr. The solution wasevaporated under reduced pressure to obtain the crude product (51.5 mg)as a yellow solid. The sample was dissolved in MeOH 1 mL and purified byMDAP (method B). The solvent was dried down to give 37 mg of a yellowoil. The oil was dried in the vacuum oven overnight. The solid (28 mg)was then was suspended in EtOH:ACN 1:1 2 mL and 80 μL of HCl in Et₂O 1 Mwere poured in. The volatiles were removed under vacuo to obtain thetitle compound (35 mg) as a yellow solid. LCMS (System J): t_(RET)=1.02min; MH⁺ 508.

Example 310: (S)-cyclopentyl2-(((1-(azetidin-3-ylmethyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-4-methylpentanoate

A round bottom flask was charged with (S)-tert-butyl3-((5-(((1-(cyclopentyloxy)-4-methyl-1-oxopentan-2-yl)amino)methyl)-2-(5-methyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-1-yl)methyl)azetidine-1-carboxylate(Intermediate 270, 60 mg, 0.099 mmol), 1,4-dioxane (10 mL) and TFA (0.5mL, 6.49 mmol). The vessel was capped and stirred at room temperaturefor 2 days. The volatiles were removed in vacuo to give a yellow oil.The oil was dissolved in DMSO (1 mL) and purified MDAP (Method B). Thesolvent was evaporated in vacuo to give the title compound (34 mg, 0.067mmol, 67.9% yield). LCMS (System B): t_(RET)=0.93 min; MH⁺ 506.

Example 311: (S)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((R)-piperidin-3-ylmethyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-methylbutanoate

A solution of 5M HCl in IPA (10 mL, 50.0 mmol) was added to(S)-tert-butyl3-((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-5-((((S)-1-isopropoxy-3-methyl-1-oxobutan-2-yl)amino)methyl)-1H-benzo[d]imidazol-1-yl)methyl)piperidine-1-carboxylate(180 mg, 0.296 mmol) (for an example preparation see Intermediate 272)and the resulting solution stirred for 1 h and stirring stopped over theweekend. The reaction mixture was evaporated in vacuo to a brown gum,dissolved in MeOH and loaded on to a 5 g SCX cartridge. The cartridgewas eluted with MeOH (50 mL) followed by 2 M methanolic ammonia (25 mL).The basic fractions were evaporated in vacuo to dryness and purified byMDAP (N31339-85-1, Formic method A). The clean, product containingfractions had 2 M aq. HCl (5 mL) added and were evaporated in vacuo to awhite solid. N31339-85-A1 was dissolved in MeOH and loaded on to a 5 gSCX cartridge. The cartridge was eluted with MeOH (20 mL), followed by2M methanolic ammonia (15 mL). The basic fractions were evaporated invacuo to give the title compound as a white solid. The total yield ofthe reaction was 67%. LCMS (System C): t_(RET)=0.48 min, MH⁺=508.

Example 312: (S)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((R)-piperidin-3-ylmethyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-methylbutanoate,dihydrochloride

(S)-isopropyl2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((R)-piperidin-3-ylmethyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-methylbutanoate(see for a preparation Example 311) (96 mg, 0.189 mmol) was dissolved inDCM (2 mL) and 1M HCl in diethyl ether (0.5 mL, 0.500 mmol) added withstirring. The resulting suspension was evaporated in vacuo to give thetitle compound as a white solid. The total yield of the reaction was97%. LCMS (System C): t_(RET)=0.48 min, MH⁺=508.

The following Example was prepared in a similar manner to Example 17:

Example 313: (2S,3R)-2-((2-(2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)ethyl)amino)-3-hydrontbutanoic acid hydrochloride (prepared from:Example 290) System D, 0.56 min, MH⁺ = 483, Yield 10 mg, 38%.

The following Examples were prepared in a similar manner to Example 95:

Example 314: (2R,3S)-isopropyl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Intermediate 116 andIntermediate 287) System E, 3.07 min, MH⁺ = 511, Yield 78 mg, 11%.

Example 315: (2R,3S)-isopropyl 2-(((1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Intermediate 224 andIntermediate 287) System E, 3.15 min, MH⁺ = 515, Yield 85 mg, 30%.

Example 316: (2S,3S)-isopropyl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Intermediate 116 andIntermediate 288) System D, 0.84 min, MH⁺ = 511, Yield 61.8 mg, 56%.

Example 317: (2R,3R)-isopropyl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Intermediate 116 andIntermediate 289) System D, 0.85 min, MH⁺ = 511, Yield 78.3 mg, 71%.

Example 318: Isopropyl 2-(((1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3-hydroxybutanoate (prepared from: Intermediate 224 andIntermediate 290) System F, 1.40 min, MH⁺ = 515, Yield 480 mg, 35%.

Example 319: Isopropyl 2-(((2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)methyl)amino)-3- hydroxybutanoate(prepared from: Intermediate 116 and Intermediate 290) System F, 1.33min, MH⁺ = 511, Yield 449 mg, 33%.

Example 320: (2S,3R)-tert-butyl2-((2-(1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)ethyl)amino)-3-hydroxybutanoate

To a solution of5-(1-(1,3-dimethoxypropan-2-yl)-5-(oxiran-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridin-2(1H)-one(for a preparation see Intermediate 291, 500 mg, 1.304 mmol) in THF (13mL) was added BF₃.OEt₂ (0.165 mL, 1.304 mmol) at 0° C. and the solutionwas stirred for 15 min. Then triethylamine (0.545 mL, 3.91 mmol) wasadded to the reaction mixture followed by (2S,3R)-tert-butyl2-amino-3-hydroxybutanoate hydrochloride (304 mg, 1.434 mmol) and TsOH(248 mg, 1.304 mmol) and reaction mixture was stirred overnight at roomtemp. Then sodium triacetoxyborohydride (276 mg, 1.304 mmol) was addedand stirring continued for another 12 hours. Sat. NaHCO₃(20 mL) wasadded to the reaction mixture and extracted with ethyl acetate (30mL×2). The combined organics were washed with brine (20 mL) dried(Na₂SO₄), filtered and concentrated to get the crude product. The crudeproduct was purified by preparative HPLC using a kinetexc8 (150*30) mm5μ column, eluting with 5 Mm Ammonium bicarbonate(Aq)/Acetonitrile. Theappropriate fractions were lyophilized to get the title compound(2S,3R)-tert-butyl2-((2-(1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)ethyl)amino)-3-hydroxybutanoate(67 mg, 9.4% yield). LCMS (System E): t_(RET)=1.46 min; MH⁺ 543.

The following Examples were prepared in a similar manner to Example 320:

Example 321: (2S,3R)-isopropyl 2-((2-(1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)ethyl)amino)-3-hydroxybutanoate (prepared from: Intermediate 291 andIntermediate 31a) System E, 1.39 min, MH⁺ = 529, Yield 45 mg, 6.4%

Example 322: (2S,3R)-cyclobutyl 2-((2-(1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)ethyl)amino)-3-hydroxybutanoate hydroxybutanoate (prepared from:Intermediate 291 and Intermediate 32) System E, 1.50 min, MH⁺ = 541,Yield 46 mg, 6.3%

Example 323: (2S,3R)-cyclobutyl 2-((2-(2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-benzo[d]imidazol-5-yl)ethyl)amino)-3-hydroxybutanoate (prepared from: Intermediate 292 andIntermediate 32) System E, 1.33 min, MH⁺ = 537, Yield 75 mg, 14%

The following Examples were prepared in a similar manner to Example 71:

Example 324: (2S,3R)-2-((2-(1-(1,3-dimethoxypropan-2-yl)-2-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1H-benzo[d]imidazol-5-yl)ethyl)amino)-3-hydroxybutanoic acid (prepared from: Example 320)System E, 1.15 min, MH⁺ = 487, Yield 45 mg, 64%

Biological Data Time Resolved Fluorescence Resonance Energy Transfer(TR-FRET) Assay

Binding was assessed using a time resolved fluorescent resonance energytransfer binding assay. This utilises a 6 His purification tag at theN-terminal of the proteins as an epitope for an anti-6 His antibodylabeled with Europium chelate (PerkinElmer AD0111) allowing binding ofthe Europium to the proteins which acts as the donor fluorophore. Asmall molecule, high affinity binder of the bromodomains BRD2, BRD3,BRD4 and BRDT has been labeled with Alexa Fluor647 (Reference CompoundX) and this acts as the acceptor in the FRET pair.

Reference Compound X:4-((Z)-3-(6-((5-(2-((4S)-6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)acetamido)pentyl)amino)-6-oxohexyl)-2-((2E,4E)-5-(3,3-dimethyl-5-sulfo-1-(4-sulfobutyl)-3H-indol-1-ium-2-yl)penta-2,4-dien-1-ylidene)-3-methyl-5-sulfoindolin-1-yl)butane-1-sulphonate)

To a solution ofN-(5-aminopentyl)-2-((4S)-6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)acetamide(for a preparation see Reference Compound J, WO2011/054848A1, 1.7 mg,3.53 μmol) in DMF (40 μl) was added a solution of AlexaFluor647-ONSu(2.16 mg, 1.966 μmol) also in DMF (100 μl). The mixture was basifiedwith DIPEA (1 μl, 5.73 μmol) and agitated overnight on a vortex mixer.The reaction mixture was evaporated to dryness. The solid was dissolvedin acetonitrile/water/acetic acid (5/4/1, <1 mL) filtered and wasapplied to a Phenomenex Jupiter C18 preparative column and eluted withthe following gradient (A=0.1% trifluoroacetic acid in water, B=0.1%TFA/90% acetonitrile/10% water): Flow rate=10 ml/min., AU=20/10 (214nm): 5-35%, t=0 min: B=5%; t=10 min: B=5%; t=100 min: B=35%; t=115 min:B=100% (Sep. grad: 0.33%/min)

The major component was eluted over the range 26-28% B but appeared tobe composed of two peaks. The middle fraction (F1.26) which shouldcontain “both” components was analysed by analytical HPLC (SpherisorbODS2, 1 to 35% over 60 min): single component eluting at 28% B.Fractions F1.25/26&27 were combined and evaporated to dryness.Transferred with DMF, evaporated to dryness, triturated with dry etherand the blue solid dried overnight at <0.2 mbar: 1.54 mg.

Analytical HPLC (Sphersisorb ODS2, 1 to 35% B over 60 min): MSM10520-1:[M+H]⁺ (obs): 661.8/− corresponding with M−29. This equates to[(M+2H)/2]⁺ for a calculated mass of 1320.984 which is M−29. This is astandard occurence with the Alexa Fluor 647 dye and represents atheoretical loss of two methylene groups under the conditions of themass spectrometer.

Assay Principle:

In the absence of a competing compound, excitation of the Europiumcauses the donor to emit at λ618 nm which excites the Alexa labelledbromodomain binding compound leading to an increased energy transferthat is measurable at λ647 nM. In the presence of a sufficientconcentration of a compound that can bind these proteins, theinteraction is disrupted leading to a quantifiable drop in fluorescentresonance energy transfer.

The binding of compounds of the invention to Bromodomains BRD2, BRD3,BRD4 and BRDT was assessed using mutated proteins to detect differentialbinding to either Binding Domain 1 (BD1) or Binding Domain 2 (BD2) onthe bromodomain. These single residue mutations in the acetyl lysinebinding pocket greatly lower the affinity of the fluoroligand (ReferenceCompound X) for the mutated domain (>1000 fold selective for thenon-mutated domain). Therefore in the final assay conditions, binding ofthe fluoroligand to the mutated domain cannot be detected andsubsequently the assay is suitable to determine the binding of compoundsto the single non-mutated bromodomain.

Protein Production:

Recombinant Human Bromodomains [(BRD2 (1-473) (Y113A) and (Y386A), BRD3(1-435) (Y73A) and (Y348A) BRD4 (1-477) (Y97A) and (Y390A) and BRDT(1-397) (Y66A) and (Y309A)] were expressed in E coli cells (in pET15bvector for BRD2/3/4 and in pET28a vector for BRDT) with a 6-His tag atthe N-terminal. The His-tagged Bromodomain pellet was resuspended in 50mM HEPES (pH7.5), 300 mM NaCl, 10 mM imidazole & 1 μl/ml proteaseinhibitor cocktail and extracted from the E coli cells using sonicationand purified using a nickel sepharose high performance column, theproteins were washed and then eluted with a linear gradient of 0-500 mMimidazole with buffer 50 mM HEPES (pH7.5), 150 mM NaCl, 500 mMimidazole, over 20 column volumes. Final purification was completed bySuperdex 200 prep grade size exclusion column. Purified protein wasstored at −80° C. in 20 mM HEPES pH 7.5 and 100 mM NaCl. Proteinidentity was confirmed by peptide mass fingerprinting and predictedmolecular weight confirmed by mass spectrometry.

Protocol for Bromodomain BRD2, 3, 4 and T, BD1+BD2 Mutant Assays:

All assay components were dissolved in buffer composition of 50 mM HEPESpH7.4, 50 mM NaCl, 5% Glycerol, 1 mM DTT and 1 mM CHAPS. The finalconcentration of bromodomain proteins were 10 nM and the Alexa Fluor647ligand was at Kd. These components were premixed and 5 μl of thisreaction mixture was added to all wells containing 50 nl of variousconcentrations of test compound or DMSO vehicle (0.5% DMSO final) inGreiner 384 well black low volume microtitre plates and incubated indark for 30 minutes at rt. 51 of detection mixture containing 1.5 nMfinal concentration anti-6His Europium chelate was added to all wellsand a further dark incubation of at least 30 minutes was performed.Plates were then read on the Envision plate reader, (⋅ex=317 nm, donor⋅em=615 nm; acceptor ⋅em=665 nm; Dichroic LANCE dual). Time resolvedfluorescent intensity measurements were made at both emissionwavelengths and the ratio of acceptor/donor was calculated and used fordata analysis. All data was normalized to the mean of 16 high (inhibitorcontrol—Example 11 of WO 2011/054846A1) and 16 low (DMSO) control wellson each plate. A four parameter curve fit of the following form was thenapplied:

y=a+((b−a)/(1+(10{circumflex over ( )}x/10{circumflex over( )}c){circumflex over ( )}d)

Where ‘a’ is the minimum, ‘b’ is the Hill slope, ‘c’ is the pIC₅₀ and‘d’ is the maximum.

Results:

All the Examples, with the exception of Examples 82, 92, 93, 271, 286and 304, were tested in the above BRD4 assay and were found to have amean pIC₅₀ in the range of 4.5 to 8.0 in the BRD4 BD1 assay and a meanpIC₅₀ in the range of 4.4 to 7.2 in the BRD4 BD2 assay, with theexception of Examples 21, 24, 48, 60 and 302 which had a pIC₅₀ of <4.3in the BRD4 BD2 assay. Example 303 was found to have a mean pIC₅₀ of 7.3in the BRD4 BD1 assay and a mean pIC₅₀ of 6.8 in the BRD4 BD2 assay.Example 68 was found to have a mean pIC₅₀ of 6.6 in the BRD4 BD1 assayand a mean pIC₅₀ of 6.2 in the BRD4 BD2 assay. Example 305 was found tohave a mean pIC₅₀ of 7.3 in the BRD4 BD1 assay and a mean pIC₅₀ of 6.4in the BRD4 BD2 assay. Example 66 was found to have a mean pIC₅₀ of 6.7in the BRD4 BD1 assay and a mean pIC₅₀ of 5.9 in the BRD4 BD2 assay. Theedisylate salt of Example 303 was found to have a mean pIC₅₀ of 7.3 inthe BRD4 BD1 assay and a mean pIC₅₀ of 6.6 in the BRD4 BD2 assay.

Measurement of LPS Induced MCP-1 Production from Human Whole Blood

Activation of monocytic cells by agonists of toll-like receptors such asbacterial lipopolysaccharide (LPS) results in production of keyinflammatory mediators including MCP-1. Such pathways are widelyconsidered to be central to the pathophysiology of a range ofauto-immune and inflammatory disorders. Blood is collected in a tubecontaining Sodium heparin (Leo Pharmaceuticals) (10 units of heparin/mLof blood). 96-well compound plates containing 1 μL test sample in 100%DMSO were prepared (two replicates on account of donor variability). 130μL of whole blood was dispensed into each well of the 96-well compoundplates and incubated for 30 min at 37° C., 5% CO₂. 10 μL oflipopolysaccharide (from Salmonella typhosa; L6386) made up in PBS (200ng/mL final assay concentration) was added to each well of the compoundplates. The plates were then placed in the humidified primary cellincubator for 18-24 hours at 37° C., 5% CO₂. 140 μL of PBS was added toall wells of the compound plates containing blood. The plates were thensealed and centrifuged for 10 mins at 2500 rpm. 25 μL of cellsupernatant was placed in a 96-well MSD plate pre-coated with humanMCP-1 capture antibody. The plates were sealed and placed on a shaker at600 rpm for 1 hour (r.t). 25 μL of Anti-human MCP-1 antibody labelledwith MSD SULFO-TAG™ reagent is added to each well of the MSD plate(stock 50× was diluted 1:50 with Diluent 100, final assay concentrationis 1 μg/mL). The plates were then re-sealed and shaken for another hourbefore washing with PBS. 150 μL of 2×MSD Read Buffer T (stock 4×MSD ReadBuffer T was diluted 50:50 with de-ionised water) was then added to eachwell and the plates read on the MSD Sector Imager 6000. Concentrationresponse curves for each compound were generated from the data and anpIC₅₀ value was calculated.

Results:

All the Examples (with the exception of Examples 2-11, 13, 15-36, 38-62,65, 67, 69-79, 81-88, 90, 91, 93-94a, 95-99, 137-143, 145, 147, 150-156,184-190, 192-210, 234-247, 271, 285-286, 288, 293, 301-302, 304, 310,313 and 324) were tested in the above assay and were found to have amean pIC₅₀ in the range of 4.6 to 8.8. Example 303 had a mean pIC₅₀ of7.6. Example 68 had a mean pIC₅₀ of 5.4. Example 305 had a mean pIC₅₀ of8.0. Example 66 had a mean pIC₅₀ of 6.0. The edisylate salt of Example303 had a mean pIC₅₀ of 7.5.

These data demonstrate that bromodomain inhibitors tested in the abovewhole blood assay inhibited the production of key inflammatory mediatorMCP-1.

Hydrolysis by hCES-1

Hydrolysis of ESM-containing BET inhibitors by carboxylesterase 1 (CES1)is one aspect of delivering a targeted molecule. Rates of hydrolysis ofcertain compounds of the invention by recombinant human CES1 weredetermined using an HPLC assay. Recombinant human CES1 (Gly18-Glu563,bearing a polyhistidine tag at the C-terminus) expressed in human cellsand purified to homogeneity was obtained from Novoprotein, Summit, N.J.,USA (catalogue number C450). Reactions were run in 384 well plates at20° C. in a buffer of 50 mM sodium phosphate pH 7.5/100 mM NaCl.

Assays used a fixed concentration of test compound (50 μM) and CES1 (50nM) and a time course of the reaction was obtained by stopping samplesat increasing times by addition of formic acid to lower the pH. Stoppedsamples were subsequently analysed by HPLC to resolve product acid fromunhydrolysed ester, using a 50×2 mm C18 5 μM reversed-phase column(Phenomenex Gemini) at a flow rate of 1 ml/min using a gradient ofacetonitrile in water, containing 0.1% formic acid. Chromatography wasmonitored using absorbance at 300 nm wavelength. The % of product formedwas determined using integrated peak areas and used to determine theinitial rate of the reaction. The specific activity of the CES1 againsteach test compound under these conditions (in units of μM/min/μM) wasobtained by dividing the initial rate of the reaction by the CES1concentration.

Results:

Examples 1, 5, 13, 14, 98, 100, 102, 104, 109a, 110, 112, 116, 119, 120,124, 125, 127, 128, 136, 139, 139a/b, 140, 141, 143, 146, 147, 155, 157,159, 160, 168, 173, 181a, 184, 187, 189, 211, 219-222, 225a/b, 228,228a/b, 229a/b, 230, 250, 258-259, 263, 268, 287, 289, 303, and 305 hadrates of hydrolysis of 0.05 to 30.92 (μM of test compound hydrolysed perminute per μM of CES1) in the above assay. Example 303 had a mean rateof hydrolysis of 0.21 (n=2) in the above assay. Example 305 had a meanrate of hydrolysis of 0.31 (n=2) (μM of test compound hydrolysed perminute per μM of CES1) in the above assay. Examples 16, 138, 156, 212,214, 216 and 162a were tested but had no detectable level of hydrolysisin this assay.

We claim:
 1. A compound of formula (Ia), or a salt thereof:

wherein R₃ is cycloalkyl, hetercycloalkyl, or the group—CHR₅(CH₂)_(a)R₆; R₅ is hydrogen, C₁₋₃alkyl, or C₁₋₃alkoxy; R₆ iscycloalkyl or heterocycloalkyl, wherein said cycloalkyl orheterocycloalkyl may be optionally substituted with one or twosubstituents selected from the group consisting of C₁₋₃alkyl,C₁₋₃alkoxy, halo, —CH₂OH, —COOH, and —COCH₃; R₇ is hydrogen, C₁₋₆alkyl,—(CH₂)_(g)cycloalkyl, —(CH₂)_(h)heterocycloalkyl, or —CR₁₃R₁₄R₁₅; R₈ ishydrogen, C₁₋₆alkyl, cycloalkyl or heterocycloalkyl, wherein saidC₁₋₆alkyl is optionally substituted with C₁₋₃alkoxy; R₁₃ is hydrogen,hydroxyl, —CH₂OR₁₈, halo, —COOH, —CONH₂, 1H-imidazol-4-yl, —SH, —SeH,C₁₋₃alkyl, C₁₋₃alkoxy, phenyl, or 4-hydroxyphenyl wherein said C₁₋₃alkylor C₁₋₃alkoxy may be optionally substituted with halo, hydroxyl,—NHC(═NH₂)NH₂, —NH₂, —COOH, —CONH₂, or —SCH₃; R₁₄, R₁₅ and R₁₈ are eachindependently hydrogen or C₁₋₃alkyl; and a, g and h are eachindependently 0, 1 or
 2. 2. The compound according to claim 1, which isin the form of a pharmaceutically acceptable salt.
 3. A pharmaceuticalcomposition comprising the compound according to claim 2, and one ormore pharmaceutically acceptable excipients.
 4. A method of treating anautoimmune or inflammatory disease, which comprises administering to ahuman subject in need thereof, a therapeutically effective amount of thecompound according to claim
 2. 5. A method of ameliorating orstabilizing rheumatoid arthritis and/or reducing or eliminatingrheumatoid arthritis symptoms, which method comprises administering to ahuman subject in need thereof, a therapeutically effective amount of thecompound according to claim 2.